ANNUAL REPORT 2000 - 2002
Institute of Plant Biochemistry A Leibniz Institute Weinberg 3 06120 Halle (Saale) Germany Phone: +49 (0) 3 45 - 55 82 11 10 Fax: +49 (0) 3 45 - 55 82 11 09 Email: [email protected] www.ipb-halle.de Table of Contents
Presentation of the Institute 4 Departmental Organization 7 Department: Stress and Developmental Biology 45 Head: Prof. Dierk Scheel Board of Directors, Foundation Council, Scientific Advisory Board 8 Research Groups: Signal Perception in Plant-Pathogen Interactions 46 Scientific Institute Council, Persons with Special Head: Thorsten Nürnberger Responsibilities, Personnel Committee 9 Cellular Signaling 48 Head: Dierk Scheel Induced Pathogen Defense 50 Department: Natural Product Biotechnology 11 Heads: Sabine Rosahl & Dierk Scheel Head: Prof. Toni M. Kutchan Metal Homeostasis 52 Research Groups: Heads: Dieter Neumann & Stephan Clemens Alkaloid Biosynthesis 12 Publications, Books and Bookchapters, Publications in press, Head: Toni M. Kutchan Patents, Doctoral Theses, Diploma Theses 54 Opium Poppy Biotechnology 14 Head: Susanne Frick Department: Secondary Metabolism 57 Plant Cell Cultures 16 Head: Prof. Dieter Strack Head: Gabriele Herrmann Alkaloid Functional Genomics 18 Research Groups: Head: Jonathan Page Molecular Physiology of Mycorrhiza 58 Mode of Action of Jasmonates 20 Head: Michael H. Walter Heads: Claus Wasternack & Otto Miersch Cell Biology of Mycorrhiza 60 Papaver-Gene Expression Analysis 22 Head: Bettina Hause Head: Jörg Ziegler Biochemistry of Mycorrhiza (since 2002) 62 Publications, Books and Book Chapters, Publications in press, Head: Willibald Schliemann Patents, Doctoral Theses, Diploma Theses 24 Glycosyltransferases 64 Head: Thomas Vogt Biochemistry of Betalains (until 2001) 66 Hops Secondary Metabolism 26 Head: Willibald Schliemann Departments of Natural Product Biotechnology and Bioorganic Chemistry - joint project Heads: Jonathan Page, Jürgen Schmidt, Frederick Stevens (until September 2002) Hydroxycinnamic Acids 68 Head: Dieter Strack Department: Bioorganic Chemistry 29 Publications, Books and Bookchapters, Publications in press, Head: Prof. Ludger Wessjohann Patents, Doctoral Theses, Diploma Theses 70
Research Groups: Department: Synthesis & Method Development 30 Heads: Ludger Wessjohann & Brunhilde Voigt Administration and Technical Services 73 Biocatalysis & Design of Ligands 32 Head: Lothar Franzen Head: Ludger Wessjohann Resources and Investments 74 Plant and Fungal Metabolites / Microanalytics 34 Heads: Norbert Arnold, Jürgen Schmidt, Ludger Wessjohann & Gernot Schneider (until June 2001) Staffing Schedule 75 Structural Analysis & Computational Chemistry 38 Use of Funds from External Sources 76 Heads: Wolfgang Brandt & Andrea Porzel Guest Researchers and Fellows 80 Publications, Books and Bookchapters, Publications in press, Patents, Doctoral Theses, Diploma Theses 40 Press and Public Relations 83 Head: Sylvia Pieplow Searching for Signals: Stress-Induced Changes in Arabidopsis Secondary Metabolite, Peptide and Map & Impressum 86 Protein Patterns (GABI) 43 Departments of Bioorganic Chemistry and Stress and Developmental Biology - joint project Heads: Stephan Clemens, Jürgen Schmidt, Ludger Wessjohann, Dierk Scheel
2 3 Presentation of the Institute
he Institute of Plant Bio - of their methodical approaches The large manifold of plant spe- biological function as well as the chemistry (IPB) in Halle was and the equipment at their dispo- cies is reflected in the enormous generation of plants with altered Tfounded on 1 January 1992 sal. This allows interdisciplinary diversity of their natural products. natural product profiles. as a non-university research insti- research using the latest chemical, This content of natural com - tute of the so-called "Blue List". In physiological, cell-biological, bio- pounds is made more complex by Molecular interactions form the 1995, the union of the Blue List chemical, molecular-biological and the change in metabolite patterns basis of cellular function. An Institutes formed the Blue List genetic methods for comprehen- during development as well as interdisciplinary analysis of these Science Association (Wis sen - sive analysis of complex subjects. when a plant is responding to its interactions is therefore of cen- schaftsgemeinschaft Blaue Liste), environment. Knowledge of the tral importance to the research which was subsequently restruc- The IPB is located on the structure and function of natural mission of the Institute of Plant tured and renamed the Leibniz Weinberg Campus, which hosts products is requisite to under- Biochemistry. The optimal adap- Association (Leibniz Ge - the natural science departments standing plant diversity, develop- tation of plants to their habitat meinschaft) in October 1997. The of the Martin Luther University, mental and adaptation processes. depends upon receptor-mediated IPB belongs to the life sciences several non-university institutes New resources can then become perception of biotic and abiotic section of the Leibniz Asso ciation. and biotechnology companies. available for innovative application environmental parameters. Ex ter - The original institute was founded Close relationships and coope- in plant production, plant protec- nal signals are evaluated, compa- as "Arbeits stelle für Bio chemie rations exist between the insti- tion, biotechnology and in the red and converted into physiolo- der Pflanzen" on 1 January 1958 tute, the university and industries. development of biologically active gical responses via altered gene by Prof. Dr. Dr. h.c. mult. Kurt Beside extensive scientific colla- compounds. Furthermore, the rea - expression patterns that are con- Mothes by order of the Ger man boration with several university lization of genome sequencing trolled by cellular and systemic Academy of Science in Berlin. In departments, the institute's de - and the growing availability of ex - signal transduction networks. The 1960 it was renamed Institute for partment heads are full profes- pressed sequence tags of various molecular basis of these proces- Biochemistry of Plants. sors at the university and, there- species is of fundamental impor- ses, receptor/ligand, en zyme/li - fore, involved in teaching and tance to functional genome analy- gand and protein/protein inter- The IPB consists of four scientific supervision of undergraduate and sis. actions, have application in the departments and the administra- graduate students. Together with development of new biologically tion and central services depart- the Institute of Plant Genetics and The comprehensive analysis of active agents. From this perspec- ment. Currently 112 employes Crop Plant Research (IPK) in plant and fungal natural products tive, the mechanisms of communi- work at the IPB paid from the Gatersleben and the Max Planck is a priority in the research mis- cation between plants and their regular budget and another 47 Institutes for Chemical Ecology in sion of the Institute of Plant symbionts and pathogens are in - fun ded by third-party funds. The Jena and of Molecular Plant Biochemistry. Structure analysis, vestigated as are biosynthetic and research profile of the institute is Physiology in Golm the IPB forms synthesis and derivatization of signal transduction pathways. unique within the German scien- the Plant Metabolism Network, natural products contribute to an Chemical struc tures of these in - tific community. The comprehen- PlantMetaNet. This network links understanding of their function teracting components are also sive analysis of natural products the plant metabolomics compe- and to an increase in their struc- modified using gene technological from plants and fungi, the investi- tence that has been developed to tural diversity. This also forms the methods, directed evolution and gation of the interaction of plants an excellent level in these four basis for investigation of their bio- chemical derivatization. The ef - with pathogens, symbionts and plant research institutes in synthesis and for discovering new fects of these changes can be mo - abiotic stresses, studies of mole- Central Germany. biologically active compounds. A nitored in model systems or with cular interactions as part of com- qualitative and quantitative analy- activity screens until a molecule plex biological processes, and sis of natural products in biologi- with the desired characteristics metabolic engineering are at the Research mission statement cal materials requires the deve- (e. g. a drug, a signal compound or center of research activities. Four thematically, methodologi- lopment of suitable analytical an enzyme) is achieved. The deve- Excellent basic research is regar- cally and organisationally overlap- methods. Subsequent identifi- lopment of new syntheses, scree- ded as the indispensable basis for ping research priorities form the cation and isolation of biosynthe- ning tests, assays and analytical the successful implementation of basis of the research mission tic enzymes can provide access to methods is supported by visuali- application-oriented research pro - state ment of the Institute of Plant the encoding genes, which in turn zation of molecular interactions jects. The institute benefits, in par- Biochemistry - plant natural pro- enables study of the regulation of via computer modelling. ticular, from the fact that the ducts, molecular interactions, the biosynthesis. The use of mu - scientific departments of the IPB information technology and meta- tants and transgenic plants ultima- A nexus of natural product re- complement each other in terms bolic engineering. tely makes possible the analysis of search and the study of molecular
4 5 Presentation of the Institute Departmental Organization
interactions is the storage and valuable chemicals, as biological evaluation of the large amount of test systems or could be of data that is generated. In particu- importance to plant breeders. lar, high through put processes used in metabolome and pro- Within four departments with teome analysis and in the produc- dis tinct, but complementary tion of combinatorial libraries research di rections and state-of- make necessary the development the-art equipment, the Institute of new methods in information of Plant Bio chemistry provides tech nology. To this end, a new optimal conditions with which junior group in information tech- to execute multidisciplinary re - nology is being established at the search in the areas of chemistry, Institute of Plant Bio chemistry. physiology, cell biology, bioche- mistry, molecular biology and Metabolic engineering is an over- genetics. The analysis of topics lapping priority in three areas of central to modern plant biology basic research - natural products, and chemistry using this wide molecular interactions and infor- array of methodologies enables mation technology. Model plants a meaningful interpretation of the are generated that have potential complex interactions in plant for various types of application. development and diversity that More specifically, de signer plants would otherwise not be pos- with tailored natural product pro- sible. The ultimate transfer of files, containing new health-pro- these results to practical appli- moting metabolites or showing cations could make ecologically improved adaptation to habitat compatible uses of plant bio- are being developed. Plants with technology a reality. < these characteristics could serve for the sustainable production of
6 7 Board of Directors, Foundation Council, Scientific Scientific Institute Council, Persons with Special Responsibilities, Advisory Board Personnel Committee
Board of Directors Scientific Institute Council Prof. Dierk Scheel Managing Director and Head of the Department Dr. Jürgen Schmidt Chairman of Stress and Developmental Biology Department Bioorganic Chemistry
Lothar Franzen Head of the Department of Dr. Otto Miersch Vice-Chairman Administration and Technical Services Department Natural Product Biotechnology
Prof. Toni M. Kutchan Head of the Department of Dr. Bettina Hause Natural Product Biotechnology Department Secondary Metabolism
Prof. Dieter Strack Head of the Department of Dr. Dieter Neumann Secondary Metabolism Department Stress and Developmental Biology
Prof. Ludger Wessjohann Head of the Department of Dr. Thorsten Nürnberger Bioorganic Chemistry Department Stress and Developmental Biology
Dr. Thomas Vogt Department Secondary Metabolism Foundation Council Dr. Brunhilde Voigt Department Bioorganic Chemistry Ministerialrat Senior Superior Counsellor Thomas Reitmann Ministry of Education and Cultural Affairs of the State of Saxony Anhalt Dr. Michael H. Walter Department Secondary Metabolism
NN Superior Counsellor Federal Ministry of Education and Research Persons with Special Responsibilities Prof. Wilhelm Boland Max Planck Institute for Chemical Ecology, Jena, Chairman of Scientific Advisory Board Dr. Gabriele Herrmann Disabled Persons’ Affairs Prof. Alfons Gierl Technical University of Munich, Vice-Chairman of Scientific Advisory Board Hans-Günter König Energy Prof. Reinhard Neubert Vice-Rector for Research and Postgraduate Students of the University of Halle Dr. Robert Kramell, Dr. Thorsten Nürnberger Radiation Protection Dr. Wolfgang Rechner Ministry of Education and Cultural Affairs of the State Saxony Anhalt Kerstin Manke Equal Opportunity Prof. Jörg Stetter Bayer AG, Leverkusen Dr. Hans-Jürgen Strunck Federal Ministry of Education and Research Sylvia Pieplow Public Relations
Scientific Advisory Board Dr. Sabine Rosahl Biological Safety Prof. Dierk Scheel, Prof. Wilhelm Boland Chairman Gene Technology (GenTG) Max Planck Institute for Chemical Ecology, Jena Prof. Claus Wasternack
Prof. Alfons Gierl Vice-Chairman Dr. Willibald Schliemannn Personal Data Privacy Technical University of Munich Prof. Thomas Boller University of Basel Dr. Hans-Jürgen Steudte Security engineer Workplace Safety Prof. Horst Kunz University of Mainz Dr. Brunhilde Voigt Eberhard Warkus Royal Veterenary and Agricultural University, Prof. Birger Lindberg MØller Copenhagen, Denmark PD Dr. habil. Personnel Committee Günter Strittmatter KWS SAAT AG, Einbeck Andrea Piskol Chairwoman Prof. Lutz F. Tietze University of Göttingen Peter Schneider Vice-Chairman Prof. Lothar Willmitzer Max Planck Institute of Molecular Plant Physiology, Potsdam-Golm Dr. Susanne Frick, Martina Lerbs, Prof. Ulrich Wobus Institute of Plant Genetics Further Members and Crop Plant Research, Gatersleben Angelika Weinel
8 9 Department: Natural Product Biotechnology Head: Prof. Toni M. Kutchan Secretary: Christine Dietel
ithin the Department of tide synthases are en - WNatural Product Biotech - coded by a multi-gene nology, the central theme of research family that has chal-cone is the analysis of the biosynthesis of synthase as a pro to type. plant natural products at the molecu- Gene fa mily evo lution in lar genetic level. Of particular inte- plants ap pears to occur rest is the isolation of genes enco- by gene duplication follo- ding enzymes and re g u latory pro- wed by nucleotide sub - teins involved in the formation of stitution that can lead to physiologically ac tive, small molecu- biochemical diversity. les derived from L-tyrosine or L- Plant polyketide syntha- tryptophan (alkaloids) and acetyl ses are presu-mably deri- Coenzyme A (polyketides). ved from a common ancestor that diverged to Alkaloids are pharmacologically perform different reac- active, nitrogen-containing, tions. The identification and char ac te - antisense or RNAi vector constructs basic compounds produced ri za tion of polyketide synthases and the influence of the transgene on in ap pro x i mately 20 % of involved in the formation a variety of metabolic pathways is deter mined by flowering plants. Each natural products should lead to a HPLC-MS. In this manner, plants with species ac cu mulates better understanding of the evolu- tailored natural product profiles can alkaloids in a u nique tion of these secondary metabolites. be generated for industrial and rese- and de fined pattern. arch use. The role of alkaloids The techniques that are used to iso- in plants has been a late and identify these genes are In addition, the signalling properties longstanding ques - wide-ranging, from enzyme purifica- of jasmonates and octadecanoids in tion, but a picture tion followed by amino acid sequen- stress-induced and developmental emerges that sup- ce determination to EST-sequencing processes con-tinues to be investiga- ports an ecochemical and macro / mi cro array analysis. ted. In particular, the spatial and tem - function for these Both plant cell cultures and native poral expression of allene oxide cycla- compounds. Al ka loid - plant material serve as a source of se alleles (a jasmonic acid biosynthe- con taining plants were enzymes and genes. The characteri- tic gene) and the physiolo gical role of also mankind's original zation of the gene products is carried jasmonic acid me ta bolites, such as “materia medica”. out after over-expression in a 12-hydroxy-jasmonate, are being Many of these plants are heterologous expression system deter mined. < still used today as sources such as bacteria, yeast, insect cells or of prescription drugs. plants. A part of gene product cha- These biosynthetic pathways racterization is the localization of the are attractive targets for mole- protein in a plant tissue or cell. To cular biology because of their role this end, antibodies are raised against in plant chemical ecology and the the he terologously-expressed bio- biotechnological potential for the synthetic proteins and immunolocali- production of commercially im - zation techniques are used to identi- portant compounds. fy the cell type in which the biosyn- thetic enzymes accumulate. This The biosynthesis of polyketides pre- potentially provides insight into the sent in medicinal plants is also under regulation of natural product biosyn- investigation. The com pounds are thesis and yields information essenti- prominent in tropical traditional al to the metabolic engineering of medicine and are used to treat a wide secondary pathways. Ulti ma te ly, the variety of ailments, with particular biosynthetic genes are transformed em pha sis on pa ra sites. Plant polyke- back into the na-tive plant as sense,
10 11 Research Group: Alkaloid Biosynthesis Head: Toni M. Kutchan
Group members The opium poppy Papaver somniferum is still today one of our most impor- proteomic analysis of P. somniferum aerial parts of the plant. latex (Decker et al. Electrophoresis 21, Exuded latex is the cyto- Kum-Boo Choi tant medicinal plants. Among the 80 alkaloids produced by this plant, three are (Humboldt Fellow since October 2002) medicinally important. These are the narcotic analgesic morphine, the analge- 3500-3516 [2000]). The cDNA was plasm and vesicles of these Torsten Grothe sic and antitussive codeine and the antitussive noscapine. The biosynthesis of amplified from P. som niferum RNA by reticulated laticifer cells. In (PhD student until April 2002) codeine and morphine is almost completely elucidated at the enzyme level. reverse transcription PCR using primers order to localize within this Robert Kramell Relatively little is understood, however, concerning the biosynthesis of nosca- based on the internal amino acid sequen- complex system the pro- (postdoctoral position since July 2001) ces. The recombinant protein was teins for which we have pine. We also understand very little of how alkaloid biosynthesis is regulated Monika Krohn expressed in Spodoptera frugiperda Sf9 isolated cDNAs, antibodies (technician since July 2001) and of the biological role of these compounds in the plant. We are systema- cells in a baculovirus expression vector. have been raised against Tobias Kurz tically isolating cDNAs that encode the unique en-zymes of alkaloid biosyn- (PhD student since April 2002) Steady state kinetic measurements with he terologously ex pressed thesis in opium poppy. These cDNAs are functionally heterologously expres- Birgit Ortel the heterologously expressed enzyme reticuline 7-O-me thyl - (technician since April 2002) sed in bacterial and insect cell cultures and characterized. The seven cDNAs and mass spectrometric analysis of the trans ferase, salutaridinol 7- Anan Ounaroon that we have isolated to date from P. somniferum will be used in in situ hybri- en zymic products suggest that the enzy- O-acetyltransferase, co dei - (PhD student until September 2002) dization and the encoded heterologous proteins in immunolocalization stu- me is capable of carry through multiple none reductase, the berbe- Khaled Sabarna (PhD student since May 2002) dies in order to identify the cellular sites of biosynthesis of some of the O-me thylations, on the isoquinoline- and rine bridge enzyme and the Marion Weid various classes of isoquinoline alkaloids (morphinan, benzo[c]phenanthridine, on the benzyl moiety of several sub - cytochrome P-450-depen- (PhD student since December 2000) phthalideisoquinoline) produced by this plant. Initial results already provide strates. The tetrahydrobenzylisoquino - dent mono oxy ge nase (S)- -1 -1 the first insight as to how biosynthesis and accumulation of the various classes lines (R)-reticuline (4.20 s mM ), (S)- N-methylcoclaurine 3'-hy - of these alkaloids are regulated in this plant. reticuline (4.50), (R)-protosinomenine droxylase. Immu no lo cali - Collaborators (1.67), and (R,S)-isoorientaline (1.44) as zation studies with each of Wanchai De-Eknamkul well as guaiacol (5.87) and isovanillic acid the antibody preparations Chulalongkorn University, Bangkok, Thailand (1.21) are O-me thylated by the enzyme is being carried out with
Tony Fist with the ratio kcat/Km shown in paren - sections of P. somniferum Tasmanian Alkaloids, Tasmania, Australia theses. A phylogenetic comparison of capsule. Initial results indi- Phil Larkins Scientific and Industrial Research Organisation Plant In recent years, we have isolated and the N-methyl moiety of (S)-reticuline to the amino acid sequence of this O- cate that multiple cell Industry, Canberra, Australia characterized cDNAs encoding several the bridge carbon C-8 of (S)-scoulerine. methyltransferase to those from 16 types are involved in al - Friedrich Lottspeich enzymes of tetrahydrobenzylisoquinoli- Specific to noscapine biosynthesis is reti- other plant species suggests that this kaloid biosynthesis in this Max Planck Institute for Biochemistry, Martinsried, Germany ne alkaloid biosynthesis from P. somnife- culine 7-O-methyltransferase. Finally, enzyme groups more closely to isoqui- plant. A heterologously ex - Werner Roos rum. The first enzyme in the biosynthetic specific to mor phine biosynthesis are noline biosynthetic O-me thy - pressed major latex protein University of Halle, Germany pathway for which we have isolated a salutaridinol 7-O-acetyltransferase and ltransferases from Coptis japonica than has been used as a latex Joachim Stöckigt cDNA is norcoclaurine 6-O-methyl- codeinone reductase, the penultimate to those from Thalictrum tuberosum. marker protein. Of the pro - University of Mainz, Germany transferase. The next is the cytochrome enzyme of the morphine pathway that teins thus far analyzed, only P-450-dependent monooxygenase (S)- reduces co deinone to codeine. It is known that morphine and other codeinone reductase can The figure shows the immunolocalization of an N-me thyl coclaurine 3'-hydroxylase. alkaloidal biosynthetic intermediates be localized to laticifer cells. These enzyme of alkaloid biosynthesis in cross-sections of These en zymes are common to the mor- Reticuline 7-O-methyltransferase con- such as dopamine accumulate in smooth results imply that intercellular transport Papaver somniferum capsule. The expression of (S)- reticuline 7-O-methyltransferase, an enzyme in - phine, noscapine and sanguinarine bio- verts reticuline to laudanine in tetrahy- vesicles within P. somniferum laticifer cells. of either intermediates or enzymes plays volved in the biosynthesis of tetrahydrobenzyliso- synthetic pathways. Specific to the san- drobenzylisoquinoline biosynthesis in In the mature plant, laticifers form a reti- a role in isoquinoline alkaloid biosynthesis quinoline alkaloids in opium poppy, occurs in the phloem of the bundle sheath (green fluorescence). guinarine pathway is the berberine P. somniferum. This new enzyme of al ka - culated system that extends throughout in P. som niferum. < The red fluorescing cells are laticifers, stained by an The pictures show incised capsules of opium bridge enzyme that oxidatively cyclizes loid biosynthesis was identified during a antibody raised against a major latex protein. poppy with protruding latex. Laticifers are the site of alkaloid accumulation in aerial plant parts. These results imply a transport of intermediates from phloem to laticifers during bio- synthesis.
12 13 Research Group: Opium Poppy Biotechnology Head: Susanne Frick
Group members Opium poppy (Papaver somniferum L.), which contains more than 80 different somatic embryogenesis from twelve cell oripavine was not always detected in B lines containing six different cDNA con- these ten transgenic plants mentioned Sandra Barth alkaloids, remains one of the most important industrial medicinal plants. (technician until December 2001) Poppy serves as a renewable resource of a number of medicinally relevant structs. After the isolation of DNA and above. One out of these ten plants show - Kathleen Gutezeit alkaloids. These include the analgesic and nar cotic drug morphine, the cough RNA, we analyzed these plants from ed an increased total amount of reticu - (technician since March 2002) suppressant codeine, as well as the muscle relaxant papaverine, the antitumo- which 150 F0 have been proven to be line instead. The pattern of the HPLC Stefanie Haase trans genic and their seeds have been via- chromatogram of the T and T plants is ric agent noscapine and the antimicrobial sanguinarine. We are developing 1 2 (PhD student since May 2002) ble. Seeds from 17 transgenic F plants almost identical. These results are a first transfor mation systems for opium poppy that will allow us: 0 Katja Kempe have not been able to germinate and 23 evidence that an alkaloid pattern in bbe (diploma student since May 2002) transgenic F plants did not contain any antisense plants is an hereditary trait. At Anja Zeuner 0 (technician) to investigate the regulation and ecological function of these alkaloids in seeds at all. the moment, we are working to confirm · the heredity of the other alkaloid pat- plants, The F0 plants were analyzed by PCR or terns observed in bbe antisense plants Collaborators by dot blots and brought to flower and and are measuring the HPLC profiles of and to alter the alkaloid metabolism in commercial poppy varieties in Tony Fist · seed set. The alkaloid pattern of the first plants containing the remaining five order to obtain varieties lacking alkaloids or with tailored alkaloid pro- Tasmanian Alkaloids, Westbury, Australia generation was determined from leaf cDNA constructs (full-length cor sense, C Phil Larkin files for industrial and pharmaceutical use. extracts by HPLC and showed altered partial cor sense, cpr sense, cyp80b1 Scientific and Industrial Research Organisation Plant Industry, Canberra, Australia alkaloid concentrations compared to sense, cyp80b1 antisense). Jürgen Schmidt During the last years, several genes from antisense orientation into explants to control plants. Molecular and chromato- Institute of Plant Biochemistry, Halle, Germany the biosynthetic pathways for reticuline, attempt to alter their alkaloid profile. graphic analysis of the F1 generation is Since we have not been able to silence sanguinarine and morphine have been Alkaloid-free plants developed in this underway for all constructs and cell ben zylisoquinoline biosynthesis in transge- cloned. Although the biosynthesis is well manner will be used to test the chemical lines. The alkaloid pattern in the second nic poppy plants containing S4S4::antiBBE understood at the enzymatic level, the ecological function of morphinan and generation is always analyzed in latex or S4S4::antiCYP80B1, we constructed molecular and biochemical mechanisms ben zo phenanthridine alkaloids in plants. and in selected plants, also in leaves and plasmids containing partial sequences that that regulate these pathways are not roots. are potentially able to trigger RNA inter- known. The goal of this project is to de - With a transgenic cell line expressing the ference in P. somniferum. Explants of D ve lop a stable transformation and rege- antisense construct of berberine bridge Latex from wild type plants of P. somni- opium poppy were transformed with neration method for opium poppy, which enzyme (BBE) we hope to reduce the ferum L. inbred parent line showed a seven different constructs: bbe RNAi, will make the metabolic engineering of metabolic flux through sanguinarine high concentration of morphine, theba - cor RNAi, cpr RNAi, cyp80b1 RNAi, 6- the above mentioned compounds possible. pathway and to enhance the concentra- ine and codeine. Another alkaloid, which omt RNAi, 7-omt RNAi and salat RNAi. Poppy seed oil finds use in chemical tion of papaverine and / or morphine is present in this extract, is oripavine. In All the explants have started to develop industry for the production of pigments instead. We are interested if a transgenic the roots of the wild type, the major calli. and lacquer, but its residual morphine cell line overexpressing codeinone re - alka-loid is the benzophenanthridine san- levels prevents more widespread appli- duc tase (COR) leads to a poppy plant, guinarine. Last year three new genes became avail - cations. As well, because opium is the which contains more morphine or where able from benzylisoquinoline biosynthe-
raw material for the illicit production of the concentration of morphine is lowered From the 150 transgenic F0 plants, 43 are sis. These encode salutaridinol 7-O-ace- heroin, cultivation of poppy is restricted. due to a possible feedback inhibition of harboring the S4S4::antiBBE construct. tyltransferase (SALAT), (S)-norcoclau - By completely suppressing morphine this pathway. We have also produced We confirmed the presence of the trans- rine 6-O-methyltransferase (6-OMT) Biosynthetic pathway from L-tyrosine to sanguinarine, papaverine and morphine in Papaver somniferum. Enzymes bio synthesis, opium poppy could become poppy transformants where we influence gene of the T1 plants with the same and (S)-reticuline 7-O-methyltransferase are highlighted in red. a "harmless" crop plant. So far, there has all cytochrome P450 enzymes of the methods described for the first gene- (7-OMT). Both methyltransferases have E been no success with breeding programs benzylisoquinoline pathways by introdu- ration. Additionally, we examined these been cloned in sense orientation in our and mutations to obtain a morphine-free cing a NADPH:cytochrome P450-oxido- plants with Southern- and Northern binary vector and have been transfor- poppy. In the best case, a reduction of reductase (CPR). Finally, we are trying to blotting. The alkaloid pattern was ana- med into opium poppy. All cultures deve- mor phine biosynthesis has been achieved. reduce or silence the complete alkaloid lyzed in latex as well as extracts from loped calli and have started to differen- The transformation of opium poppy biosynthetic pathway with a transgenic roots. tiate. < A could be an alternative to circumvent cell line containing the antisense con- these problems. struct of (S)-N-methylcoclaurine We found ten F1 plants containing hydroxy lase (CYP80B1). With a cell line S4S4::antiBBE with a different alkaloid Somatic regeneration of P. somniferum. Explants first give rise to a type I callus (A), which starts to We have used an Agrobacterium-media- overexpressing CYP80B1, we are trying pattern compared to the wild type. The differentiate after a certain time to type II callus ted approach to introduce different to stimulate all three pathways together. major alkaloids in the bbe antisense (B). After the transfer to a hormone free medium this type II callus develops small embryos (C) and cDNAs encoding enzymes of morphine The last three years we have been able plants are morphine, codeine and the ba- finally little plantlets (D). The whole regeneration process is shown in picture E. and sanguinarine biosynthesis in sense or to regenerate 190 F0 poppy plants via ine. In contrast, a peak corresponding to
14 15 Research Group: Plant Cell Cultures Head: Gabriele Herrmann
The main working task of the group is the maintenance of the plant cell cul- Group members regeneration of transfected protoplasts to plasts with a re - ture collection of our department. This collection includes about 250 different Domenika Arndt to a new suspension culture. From a duced alkaloid con- (technician) plant species of 45 different plant families. About 40 species are cultivated in number of methods we tested, the only tent (determined Ingeborg Reeh the form of suspension cultures and all others as callus cultures on various successful one was the alginate-method. spec troscopically or (technician) solid media. In figure 1 a look to a part of our suspension culture collection Fresh prepared protoplasts are mixed by HPLC analysis) is to be seen. The culture collection contains a number of plants producing with an alginate solution (a polyuronic would than be di - Collaborators interesting secondary metabolites such as alkaloids and represents the main acid from Macrocystis pyrifera) and dro- rectly the material source of biological material for coworkers and interested colleagues. In addi- plets of this mixture are solidified in for localization and Greg Pogue CaCl . Alginate clumps are then culti- characterization of Large Scale Biology, Vacaville, California, USA tion, we work in the field of alkaloid biosynthesis in the DFG project 2 vated in 24-well plates and treated with the block in the bio- Werner Roos "Functional genomics in plant cell cultures under use of viral vectors". University of Halle, Germany changing hormone media for cell division synthetic pathway. Table 1: Alkaloid content in protoplasts of and growth. After two weeks first cell For the transfec- Eschscholzia californica after elicitation with di visions are visible and after six to eight tion of protoplasts, we used two diffe- methyl jasmonate after two days. weeks, minicalli are produced. The whole rent meth ods - both PEG-mediated Molecular genetic methods should be process of regeneration takes approxi- transfection and electroporation is pos- used in the investigation of biosynthetic mately three months, which is much too sible with protoplast preparations. pathways in secondary metabolism of long. plants. One possibility to bring new To start with the search for a viral vec- genetic information into cells such as During our experiments with suspension tor, which is active in our system, we plant protoplasts could be the use of cultures of Eschscholzia californica we used a TMV-derived viral vector, which viral or bacterial vectors. Such infor- observed that also the protoplasts of should have a broad host specificity. For mation will be clones of a cDNA library, this culture can be elicitated with jasmo- a simple detection under UV light, we which should cause gain of function / nates or a yeast elicitor (table 1). This eli- cloned a Green Fluorescent Protein loss of function effects. Some cultures of citation is visible already after 24 or 48 gene into it, but we were not able to our cell culture collection contain colo- hours due to increasing amounts of san- detect any green fluorescence. The main red alkaloids visible under UV and nor- guinarine and chelirubine. With help of task now will be the search for a vector, mal light. We selected Eschscholzia cali- elicitation, transfected protoplasts can which can be transfected into Esch - fornica suspension culture because of be checked during 48 hours concerning scholzia protoplasts and expressed. < their production of a red colored mixtu- their changes in the alkaloid content. Pro - re of sanguinarine and chelirubine. In figure 2, the changes in the color of this Figure 1: A view to one part of the plant cell culture collection of the department "Natural Product suspension during one growing cycle Biotechnology": suspension cultures of different plant species. (seven days) are shown. Vectors contai- ning antisense cDNA of biosynthetic enzymes should stop alkaloid production Figure 2: The development of a suspension culture of Eschscholzia californica during one week. after successful transfection and the cells should show reduced color.
The first steps in the project were the development of a protocol for the forma- tion of protoplasts, a search for cultiva- tion or regeneration methods and a test of transfection methods for protoplasts from suspension culture. The formation of protoplasts from Eschscholzia califor- nica was done by use of cellulase and pectolyase as cell wall degrading enzymes and a purification with a Ficoll gradient (figure 3). We are now able to produce enough protoplasts of good quality and a viability of at least six to nine days. Figure 3: Protoplasts of Eschscholzia californica The next step normally should be the
16 17 Research Group: Alkaloid Functional Genomics Head: Jonathan Page
Group members We are tapping the biosynthetic potential of the plant kingdom by studying cannabis, we have constructed a tri- vitro enzymatic assay are being used to chome-specific cDNA library from puri- functional characterize these genes. Verona Dietl the metabolic pathways leading to complex natural products. Our group is (technician) using functional genomic approaches to dis-cover genes encoding enzymes fied trichome secretory cell clusters. Annegret Flier and transcription factors involved in natural product biosynthesis. This rese- More than 1.200 ESTs (expressed Stemming from the group's interest in (technician until February 2002) arch focuses on biosynthetic processes occurring in tissues or organs, such as sequence tags) from this library have the chalcone synthase superfamily of Nils Günnewich glandular trichomes that secrete natural products.We are using virus-induced been sequenced and assigned putative type III polyketide synthases, the role of (student since October 2002) gene function using bioinformatic com- these enzymes in forming medicinal plant gene silencing (VIGS) to identify genes involved in alkaloid metabolism and tri- Ursula Schäfer parisons. Through this approach we have compounds in Rheum tataricum L. (PhD student) chome development in Nicotiana benthamiana. Biochemical genomics, which identified candidate cDNA clones of (Polygonaceae), and Cassia alata L. Vincent Spelbos combines transcriptome and metabolite analysis, is being applied to uncover (student until March 2002) type III polyketide synthases, which may (Fabaceae) was studied. A new resvera- enzymes catalyzing the formation of terpenophenolic chemicals in Cannabis par ticipate in cannabinoid biosynthesis, trol-forming stilbene synthase was clo- sativa L. (hemp, marijuana) and Humulus lupulus L. (hops), see Research Group and an oxidocyclase, D9-tetrahydrocan- ned from the former (Samappito et al, in Collaborators “Hops Secondary Metabolism”. nabinolic acid synthase that is a key en- press), while a series of chalcone syntha- Valery Dolja zyme in the cannabinoid biosynthetic ses was characterized from the latter Oregon State University, Corvallis, Oregon, USA pathway. Heterologous expression and in (Samappito et al, 2002). < Jürgen Schmidt Institute of Plant Biochemistry, Halle, Germany Plants respond to virus infection by product production and storage. In the J.-Frederick Stevens silencing (turning off) viral genes and Solanaceae, they are readily targeted by Oregon State University, Oregon, USA thereby blocking viral replication. By clo- virus constructs (Figure 1). We are buil- ning plant genes into viruses, plants can ding a catalog of MYB transcription fac- be made to direct this antiviral defense tors from N. benthamiana trichomes and against their own genes, leading to a loss- testing the effect that silencing these of-function phenotype for the targeted regulatory proteins has on metabolite gene. Fast-forward genetic methods content and trichome morphology. Ex - using viruses promise to both speed up periments with known enzymes of nico- plant gene discovery and allow for the tine biosynthesis, such as putrescine-N- cloning of novel genes inaccessible to methyltransferase (PMT) and quinolate current techniques. The targets of our phospho-ribosyltransferase, have shown VIGS efforts are enzymes that gene silencing can reduce nicotine involved in tropane al- levels to about 30 % of control levels. Ba - kaloid (nicotine) biosyn- sed on these results, we are constructing thesis and transcription cDNA libraries in viral vectors for use in factors controlling meta- high-throughput VIGS approaches to bolite synthesis and accu- alkaloid biosynthesis. mulation in glandular tri- chomes of Nicotiana ben- Cannabis is grown worldwide for indu- thamiana Domin. Glan - strial purposes, yielding fibre and seeds, dular trichomes are resi- and for its content of psychoactive can- nous hairs that cover lea- nabinoids (e. g. D9-tetrahydrocannabinol, ves and flowers in many THC). The biosynthetic pathway leading plant species. Their pri- to cannabinoids is not completely under- mary function is defen- stood at the biochemical or genetic sive, although they also level. Cannabinoid biosynthesis occurs play a role in detoxifi- mainly in glandular trichomes (Figure 2) cation, and therefore they that cover female cannabis flowers at a are a major site of natural high density. Using a high-THC strain of
Figure 1: Plant virus targeting of glandular trichomes. Nicotiana benthamiana trichomes exhibit GFP expression Figure 2: Cannabinoids and terpenoids accumu- after infection with a tobacco mosaic virus containing a GFP late in glandular trichomes of Cannabis sativa. reporter gene. Photo: U.Schäfer Photo: J. Page
18 19 Research Group: Mode of Action of Jasmonates Heads: Claus Wasternack & Otto Miersch
Group members Jasmonates and their precursors, the octadecanoids, are signals in plant stress lability, an activity control of preexisting enzymes and a feed forward regulation. Carolin Delker responses and in plant development. A mechanistic analysis of the mode of (PhD student since June 2002) action of jasmonates is performed by a reverse genetics approach using the alle- This was substantiated by analysis of Tobias Kurz ne oxide cyclase (AOC)-catalyzed step in jasmonate biosynthesis. "Gain of func- Arabidopsis and mutants affected in JA (PhD student until March 2002) tion" and "Loss of function" studies with transgenic tomato plants revealed biosynthesis. Here, four different non- Claudia Kutter modulation of jasmonates and allowed to inspect the role of jasmonates in redundant AOCs are tissue-specifically (student until August 2001) active, thus allowing control of the oxyli- response to biotic and abiotic stresses as well as flower and seed development. Helmut Maucher pin signature of different organs. (postdoctoral position until March 2002) In order to use genetic approaches, functional analysis of AOC and jasmonic acid Lydia Müller (JA) is also performed in Arabidopsis thaliana. Analytics of jasmonates and other (student since October 2002) Using knockout lines of AOC1-AOC4, Jana Neumerkel oxylipins including chemical synthesis of standards and labeled substrates is an AOC1-4RNAi-lines as well as antisense (student since December 2002) essential part of this work. approaches, individual functions of Birgit Ortel AOC1-AOC4 in stress responses and (technician until March 2002) Amplification in wound-signaling by co-localization during development of Arabidopsis is Andrea Pitzschke Previous work on stress responses and of AOC, JA generation and systemin formation as under study and will allow us, to analyze (student until May 2000) well as systemin-dependent AOC expression and cloning of JA biosynthetic enzymes in JA-dependent prosystemin expression. The ampli- the mode of action of JA. Diana Schmidt Phylogenetic tree analysis for AOC's. (from (student until August 2001) barley was finished by analyses of three fication is compromised in 35S::AOCsense plans (cf. Stenzel et al. 2003a). Stenzel et al. 2003b) Ulrike Schubert 13-Lipoxygenases, three allene oxide Previously, 12-hydroxy-JA was only (student until July 2001) synthases (AOS’s) and one AOC, all of known as tuber-inducing compound in Irene Stenzel them located in chloroplasts. Since 2000, Oxylipin profiling and expression analy- Solanaceae. We could identify 12-hy - (postdoctoral position) we are working with tomato, Ara bi dop - ses in WT, 35S::AOCsense and droxy-JA and its sulfated derivative in Carola Uhlig (technician since February 2002) sis and tobacco. The first AOC was clo- 35S::AOCan ti sense lines revealed regula- A. thaliana as a signaling compound in Sabine Vorkefeld ned from tomato. This single copy gene is tion of JA bio synthesis by substrate avai- flower development. < (technician since July 2002) specifically expressed in ovules of young flowers and all vascular bundles, accom- panied by a specific pattern of various Collaborators jasmonate and octadecanoid compounds Guillermina Abdala (oxylipin signature) in distinct flower Universidad Nacional de Rio Cuarto, Argentina organs. In leaves the vascular bundle-spe- Klaus Apel cific occurrence of AOC attributes to a University of Zurich, Switzerland preferen tial generation of jasmonates in Wilhelm Boland Max Planck Institute of Chemical Ecology, Jena, Germany main veins. Based on a co-localization of Udo Conrad the AOC, the JA-generation, the location Institute of Plant Genetics and Crop Plant Research, of the wound signal systemin in vascular Gatersleben, Germany tissues and the data from various trans- Bettina Hause, Sabine Rosahl, genic tomato plants, an amplification mo - Dierk Scheel, Jürgen Schmidt Institute of Plant Biochemistry, Halle, Germany del on wound signaling is proposed. Gerd Hause University of Halle, Germany The capacity of the phloem to respond Harry Klee rapidly in wound signaling was further University of Florida, Gainesville, USA supported by detection of JA biosynthe- Occurrence of AOC protein in plastids of compa- Thomas Roitsch tic enzymes including AOC in sieve ele- nion cells (big arrows) and sieve elements (small University of Würzburg, Germany arrows)of tomato flower stalks (A, B) and petioles ments. The importance of JA in signaling John Turner (C-E). Longi tudinal sections were probed with an was strengthened by grafting experi- anti-AOC-antibody (A, C) or with the pre-im - University of East-Anglia, Norwich, UK mune serum (B). D: differential interference con- Luc Varin ments between 35S::AOCanti sense trast image of C. E: DAPI staining to visualize Concordia University, Montreal, Canada plants and wild type plants. nuclei. The sieve plate of a sieve element is marked by an asterisks (cf. Hause et al. 2003).
Immunocytological localization of AOC pro- Biosynthesis of jasmonic acid catalyzed by a lip - tein in leaves of Arabidopsis thaliana. (A) oxygenase (LOX), an allene oxide synthase (AOS), preimmune serum,(B) location of AOC in chlo- an allene oxide cyclase (AOC), an OPDA reduc - roplasts (Stenzel et al. 2003b) tase (OPR3) and b-oxidative steps.
20 21 Research Group: Papaver-Gene Expression Analysis Head: Jörg Ziegler
Group members Poppies of the genus Papaver produce a large variety of benzylisoquinoline in the data-bases. The largest groups of The expression of these cDNAs in four Andreas Gesell alkaloids. Some of them are of pharmaceutical impor-tance such as the analge- cDNAs coding for proteins with known Papaver species differing in their ability to (PhD student since July 2002) sic morphine, the antitussive noscapine or the vasodilator papaverine. The bio- function are invol-ved in tran s criptional perform the last steps in the biosynthesis Silvia Wegener synthesis to (S)-reticuline, the cen tral intermediate to all monomeric benzyli- and trans lational control, in responses to of morphine was examined and correla- (technician) soquinoline alkaloids is well understood on the molecular level, knowledge on stress, and in redox control. A nother ted with the occurrence of morphine in the later steps, which lead to the diversity of this class of compounds, is still highly re presented group codes for the respective alkaloid profiles. By combi- proteins participating in meta bolism, nation of all possible datasets, the num- Collaborators incomplete. Similarly, the regulatory steps leading to the accumulation of these mainly pri mary metabolism. To 20 ber of cDNAs possibly responsible for Birgit Dräger substances are unknown. To approach cDNA clones coding for the enzymes of sequences, a role in secondary metabo- the accumulation of morphine could be University of Halle, Germany these biosynthesis processes, we make use of the close genetic relationship, but lism could be ascribed. Five sequences reduced to 39 candidates, most of them the diversity in the alkaloid pro-file, between Papaver species or varieties, code for proteins with known function coding for unidentified proteins. Further respectively. We examine and correlate the gene expression profiles on EST- in the benzyl isoquinoline pathway, and comparisons are in progress to decrease arrays (expressed sequence tag) with specific alkaloid profiles. By the combi- for one cDNA, showing high homology the number of cDNAs far enough, that a nation of many different datasets of alkaloid profile-gene expression corre- to an enzyme involved in another alkalo- functional characterization is feasible. lations, we want to reduce the number of candidate cDNAs to a manageable id pathway that does not occur in number to start their functional characterization. Papaver, its possible role in the Another project uses the cDNA-AFLP benzylisoquino line pathway is currently technique to isolate cDNAs differentially Interloping diagram of the number of genes diffe- under investigation. The EST-sequencing expressed dependent on an alkaloid pro- rentially expressed between P. bracteatum and P. somniferum grown in the field (red circle), P. brac- project still continues, but additionally, to file. In this approach, we compare a wild teatum and P. somniferum grown in the greenhouse access cDNAs implicated in benzyliso- type P. somniferum plant with a mutant (blue circle) and P. bracteatum and P. somniferum Noscapine (green circle). The number in the over- Currently, more than 70 different poppy duction was developed and the esta- quinoline biosynthesis at a higher fre- plant accumulating thebaine, which is lapping areas indicates the number of genes that species belonging to the genus Papaver blishment of microarray technology has quency, the construction of a P450- situated four steps upstream of morphine show differential expression in the combination of the respective comparisons. have been described. Roughly, they are started. These methods are sensitive monoxygenase specific EST-collection in the biosynthetic pathway. More than able to synthesize about 2.500 different enough to record the alkaloid profile and has been initiated. These enzymes play a 100 differentially expressed fragments benzylisoquinolines, which can be grouped the gene expression pattern from one major role in the modification of the were found. Their specificity for the mor- into nine classes. The profiles of benzyl- individual plant. For 60 Papaver species benzylisoquinoline core structures lea- phine-free phenotype is currently being isoquinolines produced by the plants are and ten varieties and mutants of the ding to the high structural diversity. examined by macroarray analysis. < species-specific, however they are also opium poppy Papaver somniferum, the dependent on growth conditions. The main alkaloids could be identified by same holds true for gene expression. HPLC. For low-abundant and not yet This variability requires sensitive methods identified alkaloids, LC-MS analysis is in to record all needed parameters in one progress. As probes for the arrays, we individual plant. HPLC methods were use PCR fragments derived from an EST employed to detect the main com- project of P. somniferum stems. Among pounds of poppy alkaloids and LC-MS all Papaver species, this plant synthesizes coupling will be used for the low-abun- the largest number of different benzyli- dance compounds. For gene expression soquinolines and the stem has been analysis, a protocol for macroarray pro- shown to possess the highest biosynthe- tic activity. Up to now, we se - quenced more than 2.000 ESTs and ob - tained 1.100 uni- que sequences. A - bout 40 % ei ther code for proteins with unknown func - tion or have no ho - mology to entries
Functional classification of P. somniferum stem ESTs. Abbreviations: PS: photosynthesis, MLP: major latex proteins, protein syn/deg: pro- tein synthesis and degradation, G-prot: G-proteins, AA: amino acids, alk: alkaloids, phenyl: phenylpropanoids, isopr: isoprenoids.
22 23 Publications, Books and Book Chapters, In press, Patents, Doctoral Theses, Diploma Theses
Publications 53, 555-564 (2000). Kutchan, T. M. Aromatic and pyrone polyketides syn- Pitzschke, Andrea: Funktionelle Analyse einer Allen - Abdala, G., Castro, G., Miersch, O. & Pierce, D. Changes Kramell, R., Miersch, O., Atzorn, R., Parthier, B. & Weichert, H., Kolbe, A., Kraus, A., Wasternack, C. & Kutchan, T. M., Zenk, M. H. & Grothe, T. Salutaridinol 7- oxidcylase-cDNA durch homologe Transformation in in jasmonate and gibberellin levels during development Wasternack, C. Octadecanoid-derived alteration of Feussner, I. Metabolic profiling of oxylipins in germina- thesized by a stilbene synthase from Rheum tataricum. O-acetyltransferase and derivatives thereof in the Tomate. University of Halle-Wittenberg, Department of potato plants (Solanum tuberosum). Plant Growth gene expression and the 'oxylipin signature' in stressed ting cucumber seedlings - lipoxygenase-dependent Phytochemistry 62, 313-323 (2003). names of Institut für Pflanzenbiochemie and Meinhart of Biochemistry / Biotechnology, 5/5/2000. Reg. 36, 121-126 (2002). barley leaves - implications for different signalling path- degradation of triacylglycerols and biosynthesis of H. Zenk. patent application No. PCT/WO 02/101052 Rüder, Constantin: Untersuchungen zu Interaktions - ways. Plant Physiol. 123, 177-186 (2000). volatile aldehydes. Planta 215, 612-619 (2002). Stenzel, I., Hause, B., Maucher, H., Pitzschke, A., Miersch, A2 (2002). partnern Jasmonat-induzierter Proteine mit Hilfe des Bachmann, A., Hause, B., Maucher, H., Garbe, E., Vörös, O., Ziegler, J., Ryan, C. & Wasternack, C. Allene oxide May, C., Kindl, H., Rentz, A. & Feußner, I. The b-barrel Hefedihybridsystems. University of Halle-Wittenberg, K., Weichert, H., Wasternack, C. & Feussner, I. Kutchan, T. M. Ecological arsenal and developmental Weichert, H., Kolbe, A., Wasternack, C. & Feussner, I. cyclase dependence of the wound response and vascu- structure of lipid body lipoxygenase. PCT/WO Department of Biochemistry / Biotechnology, 19/9/2000. Jasmonate-induced lipid peroxidation in barley leaves dispatcher - The paradigm of secondary metabolism. Formation of 4-hydroxy-2-alkenals in barley leaves. lar bundle specific generation of jasmonate - 0129227 (2001). Schilling, Stephan: Isolierung und Charakterisierung von initiated by distinct 13-LOX forms of the chloroplast. Plant Physiol. 125, 58-60 (2001). Biochem. Soc. Trans 28, 850-853 (2000). Amplification in wound-signalling. The Plant J. 33, 577- Glutaminyl-Cyclase aus tierischem und pflanzlichem Biol. Chem. 383, 1645-1657 (2002). 589 (2003a). Ziegler, J., Stenzel, I., Hause, B. & Wasternack, C. Material. University of Halle-Wittenberg, Department Maucher, H., Hause, B., Feussner, I., Ziegler, J. & Weichert, H., Kolbe, A., Wasternack, C. & Feussner, I. Allenoxidcyclasegen und dessen Verwendung zum of Biochemistry / Biotechnology, 18/8/2000. Berger, S., Weichert, H., Porzel, A., Wasternack, C., Wasternack, C. Allene oxide synthases of barley Formation of 4-hydroxy-1-alkenals in barley leaves. Stenzel, I., Hause, B., Miersch, O., Kurz, T., Maucher, H., Herstellen von Jasmonsäure. German patent Kühn, H. & Feussner, I. Enzymatic and non-enzymatic (Hordeum vulgare cv. Salome) - tissue specific regula- Biochem. Soc. Trans. 28, 850-851 (2001). Weichert, H., Ziegler, J., Feussner, I. & Wasternack, C. 10004468.9 (2000). Schmidt, Diana: Analyse zur Regulation der Allenoxid- lipid peroxidation in leaf development. Biochim. tion in seedling development. Plant J. 21, 199-213 Jasmonate biosynthesis by substrate availability and cyclase-Promotoren aus Arabidopsis thaliana mittels Biophys. Acta 1533, 266-276 (2001). (2000). Ziegler, J., Keinänen, M. & Baldwin, I.T. Herbivore-indu- the allene oxide cyclase family of Arabidopsis thaliana. Ziegler, J., Stenzel, I., Hause, B. & Wasternack, C. transgener Ansätze. University of Halle-Wittenberg, ced allene oxide synthase transcripts and jasmonic Plant Mol. Biol. 51, 895-911 (2003b). Allenoxidcyclasegen und dessen Verwendung zum Department of Biochemistry / Biotechnology, 23/8/2001. Chaissaigne, H., Vacchina, V., Kutchan, T. M. & Zenk, M. H. Miersch, O. & Wasternack, C. Octadecanoid and jas- acid in Nicotiana attenuata. Phytochemistry 58, 729- Herstellen von Jasmonsäure. Japanese patent applica- Identification of phytochelatin-related peptides in monate signaling in tomato leaves (Lycopersicon escu- 738 (2001). Stenzel, I., Ziehte, K., Schurath, J., Hertel, S. C., Bosse, D. tion based on PCT/EP O1/01148, No. 100102978 Spelbos, Vincent: Prenyltransferasen in Humulus lupu- maize seedlings exposed to cadmium and obtained lentum Mill.): Endogenous jasmonates do not induce & Köck, M. Differential expression of PSI14, a phospha- (2001). lus. University of Utrecht, November 2002. enzymatically in vitro. Phytochemistry 56, 657-668 jasmonate biosynthesis. Biol. Chem. 381, 715-722 Ziegler, J., Stenzel, I., Hause, B., Maucher, H., Hamberg, tase gene family, in response to phosphate availability, (2001). (2000). M., Grimm, M., Ganal, M. & Wasternack, C. Molecular pathogen infection and during development. Physiol. Ziegler, J., Stenzel, I., Hause, B. & Wasternack, C. Schubert, Ulrike: Untersuchungen zur funktionellen cloning of allene oxide cyclase: The enzyme establis- Plant. (2003c). Allenoxidcyclasegen und dessen Verwendung zum Analyse von Allenoxidcyclase mittels Sense- und De-Eknamkul, W., Suttipanta, N. & Kutchan, T. M. Nibbe, M., Hilpert, B., Wasternack, C., Miersch, O. & hing the stereochemistry of octadecanoids and jasmo- Her stellen von Jasmonsäure. US patent application Antisense-Ansätzen. University of Halle-Witten berg, Purification and characterization of deacetylipecoside Apel, K. Cell death and salicylate- and jasmonate- nates. J. Biol. Chem. 275, 19132-19138 (2000). Vigliocco, A., Bonamico, M. B., Alemano, S., Miersch, O. (application number pending) based on Department of Biochemistry / Biotechnology, synthase from Alangium lamarckii Thw. Phytochemistry dependent stress responses in Arabidopsis are con- & Abdala, G. Activation of jasmonic acid production in PCT/EPO1/01148 (2002). 28/6/2001. < 55, 177-181 (2000). trolled by single cet genes. Planta 216, 120-128 (2002). Zea mays L. infected by the maize rough dwarf virus- Books and Book Chapters Río Cuarto. Reversions of symptoms by salicylic acid. Ellis, C., Karafyllidis, I., Wasternack, C. & Turner, J. G. The Oven, M., Grill, E., Golan-Goldhirsh, A., Kutchan, T. M. & Kutchan, T. M. Sequence-based approaches to alkaloid Biocell 26 (3), 369-374 (2002). Arabidopsis mutant cev1 links cell wall signaling to jas- Zenk, M. H. Increase of free cysteine and citric acid in gene identification. In: Phytochemistry in the Genomics Doctoral Theses monate and ethylene responses. Plant Cell 14, 1557- plant cells exposed to cobalt ions. Phytochemistry 60, and Postgenomics Era. Recent Advances in Balkenhohl, Thomas: Abbau von Speichertriglyceriden 1566 (2002). 467-474 (2002). Phytochemistry, Vol. 36. (Romeo, J.T. & Dixon, R. A., eds.) Books and Book Chapters in press in keimenden Samen der Gurke (Cucumis sativus). Pergamon Elsevier Science Ltd. Kidlington, Oxford, pp. Stenzel, I., Maucher, H., Hornung, E., Wasternack, C. & University of Halle-Wittenberg, Faculty of Feussner, I., Kühn, H. & Wasternack, C. The lipoxygena- Oven, M., Page, J. E., Zenk, M. H. & Kutchan, T. M. 163-178 (2002). Feussner, I. Transcriptional activation of jasmonate bio- Mathematics, Natural Sciences and Technology, se dependent degradation of storage lipids. Trends Molecular characterization of the homo-phytochelatin synthesis enzymes is not reflected at protein level. In: February 2000. Plant Sci. 6, 268-273 (2001). synthase of soybean Glycine max. J. Biol. Chem. 277, Kutchan, T. M. & Schröder, J. Selected cell cultures and Advanced Research on Plant Lipids. (Murata, N., 4747-4754 (2002). induction methods for cloning and assaying cytochro- Yamada, M., Nishida, I., Okuyama, H., Sekuja, J. & Fong-Chin Huang: Molecular cloning and heterologous Feussner, I. & Wasternack, C. The lipoxygenase pathway. mes P-450 in alkaloid pathways. In: Cytochrome P450 Haijime, W., eds.) Kluwer Academic Publishers, expression of Papaver somniferum cytochrome P450 Annu. Rev. Plant Biol. 53, 275-297 (2002). Oven, M., Raith, K., Neubert, R. H. H., Kutchan, T. M. & Part C. Methods Enzymol. 357 (Johnson, E.F., ed.) Dordrecht, pp. 267-270 (2003). genes involved in secondary metabolism. University of Zenk, M. H. Homophytochelatins are synthesized in Academic Press, Amsterdam, Boston London, New Munich, Department of Chemistry and Pharmacy, Frick, S., Ounaroon, A. & Kutchan, T. M. Combinatorial response to cadmium in azuki beans. Plant Physiol. 126, York, Paris, San Franciso, San Diego, Oxford, pp. 370- Stumpe, M., Stenzel, I., Weichert, H., Hause, B. & 15/5/2000. biochemistry in plants: the case of O-methyltransfera- 1275-1280 (2001). 381 (2002). Feussner, I. The lipoxygenase pathway in mycorrhizal ses. Phytochemistry 56, 1-4 (2001). roots of Medicago truncatula. In: Advanced Research Grothe, Torsten: Untersuchungen zur Morphin - Samappito, S., Page, J. E., Schmidt, J., De-Eknamkul, W. & Scheel, D. & Wasternack, C. (eds.) Plant Signal on Plant Lipids. (Murata, N.,Yamada, M., Nishida, I., biosynthese im Schlafmohn: Klonierung, heterologe Grothe, T., Lenz, R. & Kutchan, T. M. Molecular charac - Kutchan, T. M. Molecular characterization of root-spe- Transduction. Oxford University Press, Oxford (2002). Okuyama, H., Sekuja, J. & Haijime, W., eds.) Kluwer Expression und Charakterisierung der Salutaridinol-7- terization of the salutaridinol 7-O-acetyltransferase cific chalcone synthases from Cassia alata. Planta 216, Academic Publishers, Dordrecht. O-Acetyltransferase sowie Reinigung der Thebain- involved in morphine biosynthesis in opium poppy 64-71 (2002). Scheel, D. & Wasternack, C. Signal transduction in Synthase aus dem Milchsaft von Papaver somniferum Papaver somniferum. J. Biol. Chem. 276, 30717-30723 plants: cross-talk with the environment. In: Plant Signal Wasternack, C. & Abel, S. Plant hormones. In: L. University of Halle-Wittenberg, Faculty of Mathe - (2001). Schilling, S., Hoffmann, T., Wermann, M., Heiser, U., Transduction. (Scheel, D. & Wasternack, C., eds.) Molecular Plant Physiology. chapter 15 (Sharma, R., matics, Natural Sciences and Technology, 25/4/2002. Wasternack, C. & Demuth, H.-U. Continuous spectro- Oxford University Press, Oxford, pp. 1-5 (2002). ed.) Harword Press, Binghamton. Haider, G., von Schrader, T., Füßlein, M., Blechert, S. & metric assays for glutaminyl cyclase activity. Analytical Jennewein, Stefan: Klonierung und heterologe Kutchan, T. M. Structure-activity relationships of syn- Biochemistry 303, 49-56 (2002). Wasternack, C. Jasmonates - Biosynthesis and role in Expression von Cytochrom-P450-Enzymen der
thetic analogs of jasmonic acid and coronatine on Publications in press stress responses and developmental processes. In: NADPH:Cytochrom-P450-Reduktase, des Cytochrom b5 induction of benzo[c]phenanthridine alkaloid accumu- Schilling, S., Hoffmann, T., Rosche, F., Manhart, S., Abdala, G., Miersch, O., Kramell, R., Vigliocco, A., Programmed Cell Death and Related Processes in und der Taxadiensynthase aus Taxus chinensis. lation in Eschscholzia californica cell cultures. Biol. Wasternack, C. & Demuth, H.-U. Heterologous Agostini, E., Forchetti, G. & Alemano, S. Jasmonate and Plants. (Nooden, L.D., ed.) Academic Press Inc., New University of Munich, Department of Chemistry and Chem. 381, 741-748 (2000). expression and characterization of human glutaminyl octadecanoid occurrence in tomato hardy roots. York. Pharmacy 2000. cyclase: evidence for a disulfide bond with importance Endogenous level changes in response to NaCl. Plant Hause, B., Maier, W., Miersch, O., Kramell, R. & Strack, D. for catalytic activity. Biochemistry 41, 10849-10857 Growth Regul. (2003). Weichert, H., Maucher, H., Hornung, E., Wasternack, C. Anan Ounaroon: Molecular cloning and functional Induction of jasmonate biosynthesis in arbuscular (2002). & Feussner, I. Shift in fatty acid and oxylipin pattern of expression of three O-methyltransferases from mycorrhizal barley roots. Plant Physiol. 130, 1213-1220 Bailey, N. J. C., Oven, M., Holmes, E., Nicholson, J. K. & tomato leaves following overexpression of the allene Papaver somniferum L. University of Halle- (2002). Warzecha, H., Gerasimenko, I., Kutchan, T. M. & Zenk, M. H. Metabolomic analysis of the consequences oxide cyclase. In: Advanced Research on Plant Lipids. Wittenberg, Faculty of Mathematics, Natural Sciences Stöckigt, J. Molecular cloning and functional bacterial of cadmium exposure in Silene cucubalus cell cultures (Murata, N., Yamada, M., Nishida, I., Okuyama, H., and Technology, 11/9/2002. Hause, B., Stenzel, I., Miersch, O., Maucher, H., Kramell, expression of a plant glucosidase specifically involved via 1H NMR spectroscopy and chemometrics. Sekuja, J. & Haijime, W., eds.) Kluwer Academic R., Ziegler, J. & Wasternack, C. Tissue-specific oxylipin in alkaloid biosynthesis. Phytochemistry 54, 657-666 Phytochemistry 62, 851-858 (2003). Publishers, Dordrecht, pp. 275-278 (2003). Samappito, Supachai: Cloning and expression of poly- signature of tomato flower - The allene oxide cyclase (2000). ketide synthase genes from Cassia alata, Plumbago is highly expressed in distinct flower organs and vascu- Färber, K., Schumann, B., Miersch, O. & Roos, W. indica and Rheum tataricum. Chulangkorn University, lar bundles. Plant J. 24, 113-126 (2000). Wasternack, C. & Hause, B. Jasmonate - Signale zur Selective desensitization of jasmonate- und pH-depen- Patents Bangkok, 30/10/2002. Stressabwehr und Entwicklung in Pflanzen. Biologie in dent signalling in the induction of benzophenanthridi- Kutchan, T. M., Zenk, M. H. & Grothe, T. Salutaridinol 7- Hilpert, B., Bohlmann, H., Den Camp, R. O., Przybyla, D., unserer Zeit 30, 312-319 (2000). ne biosynthesis in cells of Eschscholzia californica. O-acetyltransferase and derivatives thereof in the Miersch, O., Buchala, A. & Apel, K. Isolation and charact- Phytochemistry 62, 491-500 (2003). names of Institut für Pflanzenbiochemie and Meinhart erization of signal transduction mutants of Arabidopsis Wasternack, C. & Hause, B. Jasmonates and octadeca- H. Zenk. European patent 01114122.3 (2001). Diploma Theses thaliana that constitutively activate the octadecanoid noids: Signals in plant stress responses and plant deve- Monostori, T., Schulze, J., Sharma, V. K., Maucher, H., Kutter, Claudia: Funktionelle Analyse der Allen - pathway and form necrotic microlesions. Plant J. 26, lopment. Progr. Nucleic Acid Res. Mol. Biol. 72, 165-221 Wasternack, C., & Hause, B. Novel plasmid vectors for Kutchan, T. M., Zenk, M. H. & Grothe, T. Salutaridinol 7- oxidcyclase in Lycopersicon esculentum Mill. 435-446 (2001). (2002). homologous transformation on barley (Hordeum vul- O-acetyltransferase and derivatives thereof in the University of Halle-Wittenberg, Department of gare L.) with the JIP23 cDNA in sense and antisense names of Institut für Pflanzenbiochemie and Meinhart Bio chemis- Huang, F.-C. & Kutchan, T. M. Distribution of morphinan Weichert, H., Kohlmann, M., Wasternack, C. & orientation. Cereal Res. H. Zenk. US patent application No. PCT/EP 02/07455 try / Bio technology, 23/9/2001. and benzo[c]phenanthridine alkaloid gene transcript Feussner, I. Lipids and signalling: oxylipins 3 - functional (2002). accumulation in Papaver somniferum. Phytochemistry aspects. Biochem. Soc. Trans. 28, 861-862 (2001). Samappito, S., Page, J. E., Schmidt, J., De-Eknamkul, W. &
24 25 Research Group: Hops Secondary Metabolism Heads: Jonathan Page, Jürgen Schmidt, Frederick Stevens (until September 2002)
Group members Hops (Humulus lupulus L., Cannabaceae) ments of Natural Product Biotechnology are the principal flavor ingredient in beer, (Jonathan Page) and Bioorganic Che - Marco Dessoy (PhD student since May 2002) contributing phytochemicals with both mistry (J. Frederick Stevens and Jürgen Verona Dietl taste (e.g. the bitter acid humulone) and Schmidt) aimed at clarifying the prenyl- (technician) 'nutraceutical' (e.g. the prenylflavonoid transferase reactions in hops was initia- Martina Lerbs xanthohumol) properties. These terpe- ted in 2002. We developed a sensitive (technician) nophenolic metabolites are of mixed mass spectrometric assay for in vitro Raik Löser biosynthetic origin, with precursors de- prenyltransferase activity and were able (research scientist until December 2001) rived from terpenoid and phenolic (poly- to detect the enzyme-mediated trans fer Vincent Spelbos (student until March 2002) ketide) pathways. Bitter acids and pre- of dimethylallyl diphosphate (DMAPP) to nylflavonoids are mainly made and sto- the aromatic rings of precursor com- red in specialized glandular trichomes, pounds. This industry-supported re - termed lupulin glands, found on hop search will continue with a biochemical cones (s. Figure). A key step in the bio- genomics project aimed at identifying synthesis of terpenophenolics is the genes encoding the enzymes of terpeno- transfer of isoprenoid unit(s) to the aro- phenolic biosynthesis. At a later stage we matic ring of the phenolic moiety by aro- hope to characterize the enzymes and matic prenyltransferase enzymes. A col- utilize them as biocatalysts in chemical laborative project between the Depart - transformations. <
Figure: A mature cone from hops (Humulus lupulus) with lupulin glands visible as yellow structures at the base of cone scales. Inset: Magnification of lupulin glands. (Photo: Annett Kohlberg)
26 27 Department: Bioorganic Chemistry Head: Prof. Ludger Wessjohann Secretary: Elisabeth Kaydamov
lants and fungi provide a rich source of the department are in tegrated, i. e. they span siderable action in the planning of the new Phighly diverse natural products and enzy- two or even three of these research groups. functional building (house R) in close coopera- mes. The department focuses on the isolation, Thus e. g. the projects on isoprenoids and pre- tion with the administration. House R is plan- characterization, and modification of the che- nyltransferases involve contributions from the ned to become the new home of our screening mical constituents, thereby trying to shed groups Biocatalysis & Computational Chemi - facilities and of the biocatalysis group. In addi- some light on their function in nature. The ana- stry, and to a minor extent from Synthesis & tion, it will contain cross-departmental installa- lytical work is backed by an extensive synthesis Method Development, Mi cro ana lytics and tions like a night lab, solvent distillation, a fer- program, designed to increase compound avai- Humulus. mentation room, and laboratories of other lability and molecular diversity by combinatori- departments. Also, in 2002 Phytobase was ini- al chemistry, method development, and de The present heads of research, Brun hilde tiated as a central information database plat- novo synthesis. Applications of this research Voigt, Andrea Porzel and Jürgen Schmidt were form for chemical constituents from plants, include the use of metabolites as lead structu- enforced by two new head scientists, the fungi, or valuable synthetic compounds. res for drugs, cosmetics, or as re search tools, mycologist Norbert Ar nold (Plant and Fungal Phytobase is planned to be the cornerstone of and the use of enzymes as screening targets, or Metabolites), who is also the new substitute our future information integration and partly as catalysts for synthesis. head of the department, and the biochemist will be made available within a larger context Wolf gang Brandt (Compu tational Che mis try). to all groups dependent on phytochemical data, In late 2000 the former department head, During the re port period, the group grew from including e. g. food industry, government and Günther Adam, and the interim head, Gernot some ten to about 35 members of about ten legislation, and research groups in the fields of Schneider, transferred the responsibility to nationalities. Nu merous guest researchers, natural products, nutrition, ecology, bioinforma- Ludger Wessjohann, who moved from the Vrije exchange students and probationers, from tics, metabolomics or pharmaceutical develop- Universiteit Amsterdam to Halle in early 2001. around the world as well as from local institu- ment. At the same time, the department translocated tes and schools, visited the department. The into house D and into the northern part of the closest relationships exist with colleagues in Despite the scientific and organizational unrest, now fully modernized house C. In parallel to Brazil, Vietnam, Hungary, and The Nether lands. and increasingly difficult access to outside the physical move, a scientific reorganization resources for phytochemical projects, a conti- was started. Four working groups were formed As part of the reorganization, valuable research nuous increase in publication output was achie- in 2001, three of them in totally new areas, and equip ment was newly in stalled, or re-installed ved from 2000 to 2002. Several diploma/M.Sc.- were established in the following year: and totally overhauled, among these three titles (one in Halle), and six PhDs were granted NMR and six mass spectrometer, including one to group members. Finally, Fred Stevens, who · Synthesis & Method Development FT-ICR-MS (v.i.), a glovebox, a synthetic robot, started his habilitation in the de part ment in Biocatalysis & Design of Ligands · a pipetting ro bot, and a computational che - 2000, in late 2002 accepted a call for a profes- · Plant and Fungal Meta bolites & mistry network. A fungal strain collection and sorship at Oregon State University in Corvallis Microanalytics a computerized chemical stockroom system (USA). < · Structural Analytics & Computational were started. Also, the department took con- Chem istry In addition, one interdepartmental group (GABI), working on the profiling of se con dary metabolites ("me ta bo lomics") from Arabi dop - sis thaliana, was continued in cooperation with the department of stress- and developmental biology (Dierk Scheel). A second interdepart- mental group (Humulus), studying hop con - stituents and secondary metabolism, was initi- ated with the department of plant biotechno- logy (Jonathan Page and Toni M. Kutchan). Finally, some members of the group remained active in Ams ter dam until 2003 with projects in total synthesis, especially towards new pro- drug concepts and terpenoid modifications.
It should be mentioned that despite this formal separation in research groups, most projects of
28 29 Research Group: Synthesis & Method Development Heads: Ludger Wessjohann & Brunhilde Voigt
Group members The targets of our synthetic efforts are natural products, their derivatives, and natural wards dipeptides and their derivatives proved tions (U-MCRs) leading to steroid cyclopep- Selective Organic Transformations product-like libraries, mostly of polyketide, isoprenoid or small peptoid structure, and to be the most successful. The reaction runs in tides. Using different reaction components, ma - Chromium(II) mediated reactions are highly John Bethke (postdoctoral position since June 2002) to a limited extent designer molecules, e. g. for pro-drug concepts. A crucial prere- environmentally benign solvent like ethanol, crocycle libraries were synthesized with ultima- chemoselective and allow transformations in Tran Van Chien quisite for an efficient access to such complex molecules is the availability of new but also reactions in water or without solvent te efficiency, i. e. in one step. These ma crocycles complex molecules without additional protec- (visiting PhD student since October 2002) methods with improved selectivity. The group developes these, based on our exper- are possible. No waste is produced but one are the first members of a class of host com- tion and deprotection. They also show un - Uwe Eichelberger tise in chromium and selenium reagents, biocatalytic methods (s. also dedicated rese- equi valent of water. pounds, where the rigid backbone and lipophilic common selectivity. We could demonstrate (postdoctoral position since July 2001) arch group) and multi component reactions. Selective reactions also offer the tools surface of two steroid moieties is connected via that the Hiyama-reaction to homoallylalcohols Dirk Michalik for creating chemical diversity from the modification of natural products. In our approach to 14-membered cyclopeptide peptide groups allowing variable diameters, con- can give allylketones, depending on the reac- (postdoctoral position until December 2001) Combinatorial approaches in liquid as well as on solid phase are used to obtain small alkaloids, MCRs were successfully applied for formations and chemical functions. Molecular tion conditions. These findings also suggest Lars Ostermann (postdoctoral position until June 2002) dedicated libraries, which help to find substances with im- the rapid, atom-economic construction of modeling showed that the ca vities of the synthe- solutions to the problem of low enantioselec- Eelco Ruijter proved biological activity profiles. However, the access to libraries of structurally linear precursors. For even larger, highly func- sized macrocycles are large enough to encapsu- tivity of the reaction with chiral ligands. The (PhD student since March 2001) complex, natural product like molecules is usually limited because of lengthy multistep tionalized macrocycles, a further extension of late small organic substrates (figure 3). results are used to further the development of Angela Schaks procedures. Multicomponent one-pot reactions, multiple catalytic systems, and self- "simple" MCRs like U-4CRs towards multiple Modification of Macrocycles an iterative process towards polyketide sub- (technician) selecting (evolutionary) procedures are possible solutions to improve the accessibili- structures. Günther Scheid ty of structurally complex enti ties. These processes can be applied in chemistry, e. g. (postdoctoral position until June 2002) Gisela Schmidt for selective separation and catalysis, or for pharmaceutical lead structure develop- Selenium compounds are very versatile rea- (technician) ment with an emphasis on molecules with anticancer, antibiotic, phytoestrogenic, or gents in natural product synthesis, unavoidable Henri Schrekker cosmetic properties. Especially macrocycles are of interest to us, because their con- for some transformations like the w-oxidation (PhD student since January 2001) formational design is poorly understood. They exhibit more flexibility than classical of terpenoids. However, they are toxic, odo- Tran Thi Phuong Thao aromatic drugs but have less entropy loss upon binding than open chain forms. rous and in some cases not as efficient as de- (PhD student since November 2001) sired. We are developing selenium reagents Mieke Toorneman with new characteristics: chiral, selectively (PhD student, based at Vrije Universiteit Amsterdam since February 1999) Figure 2:A 6-component macrocyclization reaction constructed from an interlocked 1.5xUgi-4-component-reaction with removable, or solid phase bound. A fluorous two bifunctional building blocks with compatible functional groups F (F = e.g. hydroxy-, sugar- or ester-moieties). phase selective reagent with improved selen- Mingzhao Zhu Pseudodilution is achieved by slow addition of one component. Head-head and head-tail cyclization and the two formed (PhD student since October 2001) Macrocycles functionality by the traditional total synthesis stereocenters will provide a library of eight isomers. oxide-elimination properties was successfully Friederike Ziethe Total Synthesis of Macrocycles approach is extremely wasteful in all resources: tested. The synthesis of chiral and solid phase (research scientist until December 2002) The most exciting polyketide macrocycles chemicals, manpower and time. It is only useful interlocked MCRs with bifunctional compo- Medicinally active macrocycles are commonly bound versions is under investigation. In a DFG discovered in recent years are the epothilones. for valuable compounds like epothilones. The nents is necessary (e. g. the 1.5-fold U-5CR, "decorated" with side-chains like lipids, phos- Research Focus Program we are investigating Collaborators They are antimitotic compounds with taxol- problem will potentiate if compound libraries depicted in figure 2). Hereby control of the phates, and heterocycles like sugars. We are a the chemical properties of selenocystein. like activity, which are also active against mul- will have to be designed, either for quantitative multiplication factor is crucial in order to avoid central partner in the EU-project ComBioCat, Jan Andreesen University of Halle, Germany tiple drug resistant cancer cell lines. Currently structure-activity relationship (QSAR) or evo- polymer formation. Towards this end, new which aims at the enzymatic modification of Uwe Bornscheuer these compounds are in phase II clinical trials. lutionary adaptation studies. Three problems com plex components have been developed polymer bound natural and natural-product- University of Greifswald, Germany One of the shortest routes to epothilones was will have to be solved for a sustainable route with two active attachment-points (amine, al - like macrocycles, their release, "decoration" Antonio Luiz Braga developed by us in Amsterdam. This approach to highly functionalized asymmetric macro- dehyde, isonitrile or carboxylic acid). and screening with self-selecting methods. We Federal University of Santa Maria, Brazil is continued towards new analogues, which pro - cycles and are addressed by our MCR-approach: could successfully establish a traceless linker Alexander Dömling, mise improved properties. Starting with bile acids suitable bifunctiona- for aldehydes on a polymer suitable for enzy- Wolfgang Richter, Lutz Weber MorphoChem AG Munich, Germany lized steroid components were synthesized matic reactions. This was exemplified with rifa- Sabine Flitsch 1. Rapid access to polyfunctional building blok- and two of such steroid units were cyclized via mycin, a macrocycle with sensitive enolether, University of Edinburgh, UK ks. peptide bridges by Ugi-multicomponent reac- acetal, acetate, and dienoate moieties, which Lucia Gardossi 2. The fast and efficient connection of these. could be modified and released as its antibiotic University of Trieste, Italy 3. Efficient macrocyclization strategies and derivative rifampicin (figure 4). Thomas Hjertberg, Bertil Helgee catalysts not based on the dilution principle. Chromium and Selenium Mediated Synthesis of Benzopyran Natural Products Chalmers University of Technology, Sweden We concentrated on the synthesis of natural Benzopyrans include such common plant Udo Kragl University of Rostock, Germany product-like macrocycles inspired by the secondary metabolites as coumarins, flavo - The structural formula of natural epothilone D, a drug 14-membered ansa-cyclopeptides from plants noids, anthocyanins etc. We are developing Rob Leurs, Martine Smit lead compound for cancer therapy. The different Free University of Amsterdam, The Netherlands colors signify structural elements originating from the and the bis-aryl-ether antibiotics (e. g. vanco- new synthetic routes towards these com- Graham Margetts building blocks used for the total synthesis by mycin). Another series is based on bifunctional pounds, especially to derivatives with new Polymerlabs UK Wessjohann et al. building blocks derived from plant metabolites, substitution patterns. < Karoly Micskei, Tamas Patonay The Fast Track to Macrocycles: especially terpenoids and steroids. Of the va - University of Debrecen, Hungary Multi Component Reactions (MCRs) rious multi component reactions, the Figure 3: Calculated conformation of macrocyclus XSA Figure 4: The macrocyclic antibiotic rifamycin bound via Romano Orru 192, C88H152N8O8, synthesized in one pot, shown with hydrazon linker to an enzyme penetrable polymeric bak- Free University of Amsterdam, The Netherlands A synthesis of designed macrocycles of high Ugi-four-component-reaction (U-4CR) to - glucose as a guest molecule. kbone (globe).
30 31 Research Group: Biocatalysis & Design of Ligands Head: Ludger Wessjohann
Group members The isoprenoid metabolism pathways provide insight into the predominant hair colored with henna- paste compared to pure law- Marco Dessoy mechanisms and routes, nature uses to build up carbon skeletons. (PhD student since May 2002) Understanding these, will provide new enzymes for in vitro C-C-coupling sone (figure 4). Thus, the first Michael Fulhorst reactions, e. g. biocatalysts for the production of prenylated and terpenoid severe analysis of the consti- (PhD student since January 2001) compounds, as well as new targets for inhibitors of important metabolic pro- tuents of henna was underta- Gudrun Hahn cesses in plants, most pathogenic bacteria, and many parasites. Of special inte- ken. This revealed not only (technician) the absence of lawsone in rest to us is the transfer of prenyldiphosphates onto aromatic substrates, as Andrea Köver fresh plant material, but also (PhD student since November 2001) neither the mechanism nor structural information about these mostly mem- the presence of several new Martina Lerbs brane bound enzymes is available. We hope to elucidate mechanistic and (technician) constituents, some of which Raik Löser structural details, provide better access to probes and substrates, develop proved cru cial for the dying (research scientist until December 2001) mechanism-based inhibitors and finally achieve access to a set of enzymes capability of the plant materi- Lech Luczak enabling a multitude of enzymatic C-C-coupling reactions. Figure 2: The influence of co-solvents and their concentration on al. For the first time we could (postdoctoral position since August 2002) the geranylation yield of 4-hydroxybenzoate catalyzed by ubiA-oli- prove that the liberation of Fred Stevens But also other enzymes, e. g. hydrolyases and oxidoreductases are used for goprenyltransferase from E. coli. (postdoctoral position until October 2002) dye precursors proceeds the efficient synthesis of building blocks, especially for enantio- or regioselec- Figure 3: The chemical structure of the henna-dye lawsone Svetlana Zakharova and a first chromogenic assay was deve- enzymatically, and that a complex pro- (postdoctoral position since November 2002) tive transformations. In some cases, enzymatic reactions can also be used for loped. The important factors, which dye concept is active in the plant powder. pro-drug like systems, or such systems are discovered in nature. govern the enzyme stability were identi- Apart from studying the enzymatic trans- fied, and consequently the yield of pro- formation, we also started to look at the Collaborators duct could be improved to almost 99 % conjugation of the final dye lawsone to Henna, Henna, Lawsone leaf paste water extract Jürgen Allwohn for the natural substrate, reducing at the peptides and proteins. Wella AG, Darmstadt, Germany same time the amount of enzyme requi- Han Asard Prenyltransferases and Isoprenoid the intermediates of the latter synthesis. red. Other University of Nebraska-Lincoln, USA Compounds UbiA-prenyltransferase is a membrane We could achieve the regioselective enzy- Uwe Bornscheuer University of Greifswald, Germany The synthesis of isotope labeled tenta- bound enzyme that catalyzes the oligo- Henna matic resolution of epothilone acyloin buil- Bettina Hause tive metabolites of the new non-mevalo- prenylation of 4-hydroxybenzoic acid Henna is a powder from dried leaves of ding blocks as part of our on-going effort Institute of Plant Biochemistry, Halle, Germany nate (dxp-) pathway was achieved. In a (PHB) in 3-position as part of the bio- the henna-plant Lawsonia inermis. Since towards an effective synthesis of epothilo- Lutz Heide collaboration, some compounds were synthesis of ubiquinones (figure 1). some 3000 years, henna is used for the ne derivatives (cf. research group "synthe- University of Tübingen, Germany utilized to prove for the first time, that Previously we could demonstrate the temporary dyeing of hair or skin. sis"). The best suitable lipases were identi- Udo Kragl 4-hydroxy-dimethylallyldiphosphate use of the E. coli - enzyme in vitro and 2-Hydroxynaphthoquinone (lawsone, fied. We also tested the selective enzyma- University of Rostock, Germany (4-OH-DMAPP) is an intermediate of elaborate a model of the aromatic sub- figure 3) was considered the main ingre- tic acylation/deacylation of macrocyclic Romano Orru Free University of Amsterdam, The Netherlands the new pathway in plants. A modified strate. This model was extended and dient responsible for the dyeing. How - es ter and hydroxy side-chains in solid phase Markus Pietzsch Poulter-procedure for the synthesis of improved, and for the first time also one ever, distinct differences were found in gels. < University of Halle, Germany very sensitive aldehyde-pyrophosphates for the prenyl component was developed. Kazufumi Yazaki was developed, in addition to our new Both substrate models were incorpora- University of Kyoto, Japan synthesis of organic diphosphates and ted in the first protein models of this Meinhart Zenk cyclic phosphates. Furthermore, we stu- class of transferases, based on homology University of Halle, Germany died the physico-chemical properties of calculations (cf. research group "compu- tational chemistry"), which form the basis for future verification by site direc- ted mutagenesis and mechanism-based inhibitors. The synthesis of several such inhibitory compounds was successfully started.
For the better production of prenylated hydroxybenzoates, the influence of Figure 4: Fluorescence microscopy of cross-sec - various parameters and modifiers like tioned hairs dyed by various preparations. It is cosolvents (figure 2), additives and metal clear ly visible that henna leaf-paste, still containing active enzyme, dyes hair throughout and fully, Figure 1: The enzymatic oligoprenylation (n > 1, OPP = diphosphate) of p-hydroxy benzoate (PHB). ions on the reaction was studied. where as a water extract and synthetic lawsone Improved assay-conditions were found, possess greatly reduced almost identical dyeing properties (Photo by Bettina Hause).
32 33 Research Group: Plant and Fungal Metabolites / Microanalytics Heads: Norbert Arnold, Jürgen Schmidt, Ludger Wessjohann & Gernot Schneider (until June 2001)
Group members Defined natural substances find numerous applications in society and indu- situation has stabilized, because the past cooperation was highly successful with Nguyen Hoang Anh stry, e.g. as chemical raw materials, food additives, cosmetics, in agriculture, (postdoctoral position until August 2001) and especially in medicinal chemistry where they form the basis of more than respect to our local partners, structures Torsten Blitzke one third of all currently approved drugs. The research group focuses on the or biological activities found. (postdoctoral position until May 2000) isolation and chemical characterization of pure compounds from plants and Katrin Franke fungi, which are a rich and diverse source of secondary metabolites. In addi- The Americas (postdoctoral position since September 2000) Though the political development in tion, techniques for the improved analysis and profiling of plant metabolites Gudrun Hahn Latin America is very favorable, natural (technician) are developed, especially through the application of mass spectrometry. The 6-(8''-umbelliferyl)-apigenin from Gnidia socotrana product research is increasingly ham- Tobias Herzfeld biological activities of extracts, fractions and especially pure substances are (Thymelaeceae) (PhD student until September 2002) pered by bureaucracy for both, the local Myint Myint Khine tested in bioassays. These are designed to elucidate the function of the tested (Moraceae) eleven new furanocouma- partners as well as the international (PhD student since September 2002) compounds in na ture or to screen for medicinally or otherwise useful pro- rins, especially with oxygenated geranyl ones. Together with a veterinary insti- Oil flower of Ennelophus euryandrus (Iridaceae, Christine Kuhnt perties. chains, as well as one benzofuran deriva- tute in Southern Brazil we looked at photo by Günter Gerlach), and formula of a typi- (technician) cal floral oil component with a 2-acetoxy fatty tive could be structurally elucidated by pampas plants toxic for cattle. Consti - acid residue. Monika Kummer high-resolution MS and 2D-NMR-analy- tuents, which are potentially responsible (technician) cation of approximately 150 substances. sis. With a series of new cardanols a for the observed bone deformation or Martina Lerbs Plant metabolites: (technician) Southeast Asia Besides numerous known compounds potential cancerostatic activity were even death, have been identified. Tilo Lübken HEA(N)TOS is a drug used for an effec- from ma ny classes, several new com- identified by GC-MS in Rhus thyrsiflora (PhD student since March 2001) tive detoxification treatment of drug pounds were detected. (Anacardiaceae). Moreover, besides Rather than being guided by ethnophar- Jana Mühlenberg addiction. It is based on the traditional some known piperidine alkaloids a new mocological, phytochemical or observa - (PhD student since August 2002) herbal medicine of Vietnam. The abbre- Apart HEA(N)TOS, the investigation of chlorinated amide from Aloe sabaea tive selection of plants, in a different Ernst Plaß (postdoctoral position until August 2001) viation HEA(N)TOS is derived from medicinal plants from Southeast Asia, (Aloeaceae) as well as a new 5-methyl- approach we concentrate on the tissue Lars Seipold "heat of the sun", and was originally especially Vietnam and Myanmar (Birma), chromone glycoside from Commi phora specific profiling of constituents from (PhD student since January 2000) developed by the Vietnamese herbalist is continued. A PhD-thesis on consti- socotrana (Burseraceae) could be isola- secretive plant organs. The emphasis is on Trinh Thi Thuy Tran Khuong Dan. It is now produced in tuents of Fissistigma spec. from Vietnam ted. The first finding of two coumarin- elaiophores, floral glands that produce (postdoctoral position until October 2002) an improved formula at the Institute of was concluded and numerous new flavo- flavonoid hybrid compounds from Gnidia oils as rewards for pollinators of predo- Nguyen Hong Thi Van Chemistry of the National Center for noid-terpenoid-hybrid compounds could socotrana (Thymelaeceae), representing minantly neotropic plants. The oil flower (guest scientist since April 2002) Natural Sciences and Technology in be published. a new type of compounds, was an impor- syndrome was not discovered be fore the Centris bee visiting the oil flower of Malpighia Hanoi. It is composed of 13 medicinal tant topic in the report period. A phyto- seventies. The chemical composition of emarginata. Photo by Günter Gerlach plants and natural products grown in chemical investigation of Eulophia peter- floral oils is mostly unknown; the analyti- Vietnam. As part of an UNESCO pro- sii (Orchidaceae) led to the identification cal methods for their profiling and relia- ject concerning the international scienti- of structurally known phenanthrene ble analyses were not yet established. In fic development and standardization of derivatives (cooperation with Mohamed cooperation with the Botanical Garden the anti-drug medication HEA(N)TOS, Masaoud, Sanáa). Munich-Nymphenburg (Günter Gerlach) we perform phytochemical studies of the and several international partners (e. g. constituents in close cooperation with In cooperation with Luay Ra shan (Am - Beryl B. Simpson), we gained access to Sung Tran from the Institute of Chemis - man) the trail for a chemical basis for various species of oil flower plants. The try in Hanoi. The aim of our work is the the anticancer properties of a regional analytical work included the collection of isolation and structural elucidation of plant was followed. The work on consti- the floral oils by microscopic techniques, the compounds with potential biological tuents of African species in Antidesma, development of micro-derivatization activities and the compilation of litera- which led to the discovery of com- methods, and analysis of the volatile deri- ture data on the components already pounds highly active against trypano- vatives by GC/MS. The underivatized oils known. These investigations contribute somes (Chagas' disease) was concluded. were analyzed by electrospray tandem to the botanical identification of the A new project based on plants from mass spectrometry. Structures of The traditional Vietnamese herbalist Dan and his used plant species and provide the Hea(n)tos drug preparation. Madagascar was initiated. However, as uncommon chain oxidized lipids could be necessary prerequisites for further deve- with other projects in this world region, elucidated, e. g. novel dihydroxylated fatty lopment and a future global use of Africa, Mediterranean, and Middle East the cooperation was halted for the time acids and their glycerides in the floral oil HEA(N)TOS. Until now, our group inve- In the course of a phytochemical project being. from species of the families Malpi ghi - stigated the constituents of seven of the with the Sanáa University (Yemen), a aceae and Orchidaceae. The investigation 13 components. So far, the phytochemi- series of new natural compounds were We hope to re-establish active projects, of the secretions from seven different, cal investigations resulted in the identifi- found. Thus, from Dorstenia gigas when the safety, political and financial non-related plant families supported the
34 35 Research Group: Plant and Fungal Metabolites / Microanalytics Heads: Norbert Arnold, Jürgen Schmidt, Ludger Wessjohann & Gernot Schneider (until June 2001)
Collaborators fact, that plant diversity is reflected in dif- could be isolated and their structures tabacum (Otto Miersch, Halle; Luc and depsidones from the lichen Lecidea ferent chemical compo-sitions of the flo- elucidated as cyclopentenon derivatives. Varin, Montreal). Betalains, resveratrol inops were analyzed (Siegfried Huneck). Joe Ammirati University of Washington, Seattle, USA ral oils, and that the syndrome evolved Information about their biosynthesis is gluco side, sinapic acid derivatives, flavo- Phytosterols as marker for specific mu- Marta Andriantsiferana independently. In a related project, the expected from feeding experiments with noids and other secondary metabolites tations during the embryogenesis of University of Antananarivo, Madagascar floral glands of hops (Humulus lupulus) 13C-labeled precursors. In addition, some could be also evaluated by MS/MS tech- Arabidopsis could be identified by GC-MS Helmut Besl are studied in a cooperative effort with compounds show remarkable antifungal niques (Willi bald Schliemann, Alfred (collaboration with Katrin Schrick and University of Regensburg, Germany the department of plant biotechnology activity in our bioassay. Baumert, Thomas Vogt, Dieter Strack, Gerd Jürgens, Tübingen). Manfred Binder (see separate chapter). Halle). In collaboration with the group of Clark University, Worcester, USA The fruitbodies of Cortinarius bolaris, a Lutz Heide (Tübingen) especially the LC- In 2001, the mass spectrometry facilities Joao Braga de Mello Universidade Federal de Rio Grande do Sul, Fungal metabolites: species described in the literature as ESI-selected reaction monitoring (SRM) as were improved by establishing the elec- Porto Alegre, Brazil The kingdom of fungi is composed of an poisonous, are staining yellow when bru- a sensitive and effective method for trace trospray Fourier-transform ion cyclo- Günter Gerlach estimated 106 specimen and forms on of ised or cut. The yellow stained areas analysis, has led to the identification of a tron resonance technique. This new Botanical Garden Munich, Germany the biggest group of organisms series of new aminocoumarin antibiotics of technique allows mass spectral analyses Lutz Heide, Shu-Ming Li University of Tübingen, Germany in our world. Many of them are the novobiocin-, chlorobiocin - and cou- with very high resolution and mass ac - Jochen Kopka living in symbiosis with plants mermycin type. Pro ducts of the polyketide cacy. This allowed the solution of some Max Planck Institute of Molecular Plant Physiology, (mycorrhizal fungi), others are synthesis were identified by LC-ESI-MS/MS diffi cult problems with synthetic com- +Golm, Germany pathogenic. At this time only (Toni M. Kutchan, Jonathan Page, Halle). pounds (e. g. macrocycles) and for the Toni M. Kutchan, Jonathan Page, Dierk 5 % (75.000) of all fungi are Using both, positive and nega tive ion elec- identification of natural products. < Scheel, Stephan Clemens, Dieter Strack, well described. In our research trospray, perlatolic acid derived depsides Alfred Baumert, Willibald Schliemann on fungal metabolites, we are Institute of Plant Biochemistry, Halle, Germany 7-Tesla-Fourier-transform ion cyclotron reso - Kurt Merzweiler main ly focused on compounds nance mass spectrometer (FT-ICR-MS) with an University of Halle, Germany from fruitbodies of Basidio - electrospray (ESI) ion source Luay Rashan mycetes (e. g. Hygro phorus, Applied Science University, Amman, Jordan Cor tinarius). Species in the Joachim Schröder genus Hygro phorus Sect. Cortinarius bolaris (day light) Cortinarius bolaris (UV 365 University of Freiburg, Germany nm) Oliva ce oum brini are well cha- Beryl B. Simpson University of Texas, Austin, USA racterized by a yellow reaction after tre- show a bright golden fluorescence in UV- Wolfgang Steglich ating the stem with base like potassium light. The chemical principle underlying University of Munich, Germany hydroxide. The responsible constituents these phenomena could be isolated and Tran Van Sung was characterized as a new benzofuran NRCS, Institute of Chemistry, Hanoi, Vietnam glycoside. Further research is directed at Meinhard H. Zenk the chemical constituents of Sepe doni - Biocenter Halle, Germany um (Fungi imperfecti), which live as para- sites on boletes and bolete relatives (Bo le- ta les).
Microanalytics: The coupling of HPLC and electrospray (ES) tandem mass spectrometric methods was successfully applied to the micro- analysis of a series of natural compounds in collaboration with all departments of the IPB and external groups. Thus, a LC-ESI-MS/MS method for the determi- nation of 5-methylchromone glycosides was developed and some new com- pounds of this type from Aloe species could be identified. 12-hydroxysulfonyl- oxyjasmonic acid was identified by selected reaction monitoring in Feeding experiment on Hygrophorus latitabundus Arabidopsis thaliana and Nicotiana
36 37 Research Group: Structural Analysis & Computational Chemistry Heads: Wolfgang Brandt & Andrea Porzel
Group members The research group is investigating three-dimensional molecular structures of such as SYBYL, MOE (Molecular Monika Bögel small molecules and proteins as well as reaction mechanisms in the field of Operating Environ ment) SPAR- (research scientist since August 2001) bioorganic chemistry by means of molecular modeling, semi-empirical calcu- TAN, JAGUAR and GAUSSIAN Lars Bräuer lations, nuclear magnetic resonance (NMR) and optical spectroscopy. The were installed (figure 3). (student until June 2002, afterwards PhD student) group is also respon sible for the development of a database, designed to solve Alexander Buske The first project aims at the (research scientist until December 2000) problems involved with phytochemical investigations such as fast derepli- homology modeling of aromatic Susanne Drosihn cation (Phytobase). The collected information together with data mining and prenyltransferases, an important (research scientist until February 2002) new data from the other research groups forms the basis for chemoinforma- group of enzymes of which neither Dubravko Jelic tic analyses, which will enable new insights in the biological significance of (guest scientist until March 2002) Figure 2: Constitution and configuration of two new aurones 3-D structural information nor Olaf Ludwig plant and fungal metabolites. mechanistic details of the cata- (system administrator since March 2001) test of the Federal Institute for Materials lysis mechanism were known. Most aromatic Maritta Süße Research and Testing (BAM, Berlin) for the prenyltransferases are membrane bound, like (technician) In 2000, the DFG supported project 300 MHz NMR spectrometer were equipped validation of 1H NMR spectroscopy as a reliably 4-hydroxybenzoate oligoprenyltransferase Larisa Vasilets (guest scientist until December 2002) "Conformation and structure-activity re- with new radio-frequency consoles and new quantitative analysis method. The main task (ubiA), a key enzyme in the biosynthesis lationship of brassinosteroids" was finished probe heads. Inter alia, for the first time high- of the NMR-subgroup was the structural elu- pathway of ubiquinone. It catalyzes the preny- with investigations of the side-chain confor- resolution MAS (magic angle spinning) proton cidation of natural products and synthetic lation of 4-hydroxybenzoate in the 3-position Figure 4: The most likely model of 4-hydroxyben- zoate oligoprenyltransferase with docked substra- Collaborators 13 mations of brassinosteroids in aqueous solu- spectra, deuterium decoupled C spectra and com pounds in collaboration with the other with an oligoprenyldiphosphate and is one of tes octaprenyl-diphosphate (OP-PP), 4-hydroxy- APOGEPHA GmbH tion with and without the presence of micelle DOSY (diffusion ordered spectroscopy) spec- groups of the department. As an example of the best-characterized examples, which was benzoate (4-HB) and magnesium (Mg) at the expected active site. Dresden, Germany forming agents. Using the sophisticated WET tra could be recorded. A new 400 MHz NMR a successful structural elucidation, figure 2 also available to us for experimental verifica- Horst Bögel solvent signal suppression technique (water spectrometer equipped with a four nuclei shows two new aurones (collaboration with tion (cf. biocatalysis group). By using homo - responding docking studies the structure- University of Halle, Germany suppression enhanced through T1 effects), auto-switchable probe was installed in that Fred Stevens). The constitution as well as the logy modeling and multiple alignments, activity Dieter Brömme (figure 1) The Mount Sinai School of Medicine, New York, USA spectra with sufficient signal-to-noise ratios year . Since March 2002, this spec- configuration of the double bonds could be secondary structure prediction, molecular Volker Christoffel, could be recorded, even if the solubility of trometer is operated as an open-access rou- elucidated by one- and two-dimensional dynamics simulations and energy optimiza- Barbara Spengler brassinosteroids in water is less than tine NMR for trained graduate students, post- NMR experiments and NOE investigations. tions, two first models with two possible acti- Bionorica AG, Neumarkt, Germany 0.2 mmol/l. A highly conserved solution docs and technicians of the department. In ve sites could be created and refined (figure re lation ships of a multitude of opioids could Ivo Feußner struc ture of the steroidal side-chain was preparation for the open access use, a set of In late 2001, the subgroup "computational 4). be explained and new derivatives with impro- University of Göttingen, Germany found in case of brassinolide whereas the less macro programs was developed, which allow chemistry" was started. A powerful computer- ved properties could be proposed for synthe- Susanna Fürst bioactive 24-epi-brassinolide showed diffe- the easy set-up of experiments and data pro- cluster of altogether four UNIX-workstations Other investigations concerned the analysis sis. Further more, the unusual long duration of Semmelweis University of Budapest, Hungary rent conformations dependent on the cessing. This included solvent de pend ent shim and six LINUX and WINDOWS-PCs was of structure activity relationships of epothi - action of a new class of kappa selective Lutz Heide, Shuming Li University of Tübingen, Germany medium. sets and parameter files, which were adjusted installed as basic prerequisite for the perfor- lones accompanied by conformational inve- opioids could be clarified based on mechani- Ulrike Holzgrabe at regular intervals. All users were trained in mance of molecular modeling calculations. stigations based on NMR data, conformatio- stic investigations and ab inito DFT-calcula- University of Würzburg, Germany The NMR equipment was largely modernized the operation of the NMR spectrometer and For this purpose, software program packages, nal studies of macrocycles, and calculations tions. Andris Kreicbergs and expended with the different focus of the had to pass an "NMR driving test" before on the reaction mecha- Karolinska Institute, Stockholm, Sweden department in 2001. The 500 MHz and the using the instrument unsu per vised. nism of the late enzymes In 2002, "Phytobase" was started in collabo- Volker Lipka of the non-mevalonate ration with Volkmar Vill (Hamburg) as a long- Max Planck Institute of Plant Breeding Research, Cologne, Germany Service measurements of pathway (MEP-pathway) term development project. The compound Klaus Neubert NMR spectra as well as of isoprenoid biosynthe- based reference and spectroscopy collection University of Halle, Germany opto-analytical (IR, UV, sis, especially the conver- will include biological information and will PLIVA AG CD and ORD) spectra sion of 2-methyl-D-ery - directly link all available data in an object- Zagreb, Kroatien were carried out for thritol-2,4-cyclodiphos- oriented database. Phyto base will be the cen- Dierk Scheel, Dieter Strack, Thomas Voigt, Judith Hans scientists of this and other phates. tral information system of the department Institute of Plant Biochemistry, Halle, Germany departments of the IPB. and is a cornerstone for future development Helmut Schmidhammer Since 2000, the sample Within the EU-project of the IPBs natural product research. It will be University of Innsbruck, Austria volume increased several "Opioid Treatment of available to the other departments, and parti- Sungene GmbH times. Currently some Chro nic Pain and Inflam - ally to external partners (PlantMetaNet, fun- Gatersleben, Germany 5000 spectra are recor- mation of the Loco motor ding partners) and the public. < Volkmar Vill University of Hamburg, Germany ded per annum. The NMR System", 3D-models of Meinhard H. Zenk laboratory of the IPB par- the opioid receptors have University of Halle, Germany Figure 1:The new 400 MHz NMR spectrometer used by Dr. Trinh Thi Thuy ticipated successfully in a Figure 3: Configuration of the workstations and PC cluster of the modeling been developed. Based in open-access mode national interlaboratory group. on these models and cor-
38 39 Publications, Books and Bookchapters, In press, Patents, Doctoral Theses, Diploma Theses
Publications Maier, W., Schmidt, J., Nimtz, M., Wray, V. & Strack, D. Schulz-Lang, E., Burrow, R. A., Braga, A. L., Appelt, H. Wessjohann, L. A. Synthesis of natural-product- Adam, G. 25 Jahre deutsch-vietnamesische Buske, A., Schmidt, J. & Hoffmann, P. Chemo - Gräther, O. & Schneider, B. The metabolic diversity Secondary products in mycorrhizal tobacco and R., Schneider, P. H., Silveira, C. C. & Wessjohann, L. A. based compound libraries. Curr. Opin. Chem. Biol. Zusammenarbeit auf dem Gebiet der pflanzlichen taxonomy of the tribe Antidesmeae (Eu - of plant cell and tissue cultures. Physiology, tomato roots. Phytochemistry 54, 473-479 (2000). R,R-(+)-Bis[(3-benzyloxazolan-4-yl)methyl]disulfi- 4, 303-309 (2000). Naturstoffchemie - ein Resümee. Viet Nam Info 1, phorbiaceae): antidesmone and related com- Progress in Botany 62, 266-304 (2001). de. Acta Cryst. E57, 41-42 (2001). 5-6 (2000). pounds. Phytochemistry 60, 489-496 (2002). Nguyen Thi Hoang Anh & Tran Van Sung, Some Wrenger, S., Kähne, T., Faust, J., Mrestani-Klaus, C., Holzgrabe, U., Cambareri, A., Kuhl, U., Siener, T., results on Phytochemical study of Rehmannia gluti- Shu-Ming Li, Westrich, L , Schmidt, J., Kuhnt, C. &. Fengler, A., Stöckel-Maschek, A., Lorey, S., Brandt, W., Amaral, A. C. F., Kuster, R. M., Bessa, W. d. S., Barnes, Buske, A., Schmidt, J., Porzel, A. & Adam, G. Brandt, W., Straßburger, W., Friderichs, E., nosa rhizomes. Proceeding of national conference Heide, L. Methyltransferase genes in Streptomyces Neubert, K., Ansorge, S. & Reinhold, D. R. A., Kaplan, M. A. C. & Wessjohann, L. A. Flavonoids Alkaloidal, Megastigmane and Lignane Glucosides Englberger, W., Kögel, B. & Haurand, M. on organic chemistry, 329-332 (2001). rishiriensis: new coumermycin derivatives from Downregulation of T cell activation following inhi- and other phenolics from leaves of two Marliera from Antidesma membranaceum (Euphorbiaceae). Diazabicyclononanones, a potent class of kappa gene inactivation experiments. Microbiology 148, bition of dipeptidyl peptidase IV/CD26 by the N- species (Myrtaceae). Biochemical Systematics and Eur. J. Org. Chem. 2001, 3537-3543 (2001). opioid analgesics. Il Farmaco 57, 531-534 (2002). Nguyen Thi Hoang Anh, Tran Van Sung, Porzel, A., 3317-3326 (2002). terminal part of the thromboxane A2 receptor. J Ecology 29, 653-654 (2001). Franke, K. & Wessjohann, L. Homoisoflavonoids Biol Chem. 275, 22180-22186 (2000). Cassán, F., Bottini, R., Schneider, G. & Piccoli, P. Holzgrabe, U., Friderichs, E., Englberger, W., Kögel, from Ophiopogon japonicus Ker-Gawler. Smagghe, G., Decombel, L., Carton, B., Voigt, B., Baumert, A., Mock, H.-P., Schmidt, J., Herbers, K., Azospirillum brasiliense and Azospirillum lipoferum B., Haurand, M., Strassburger, W., Brandt, W., Phytochemistry 62, 1153-1158 (2002). Adam, G. & Tirry, L. Action of brassinosteroids in Sonnewald, U. & Strack, D. Patterns of phenylpro - hydrolyse conjigates of GA20 and metabolize the Cambareri, A., Kuhl, U. & Siener, T. the cotton leafworm Spodoptera littoralis. Insect Books and Book Chapters panoids in non-inoculated and potato virus Y-inocu- resultant aglycones to GA1 in seedlings of rice Diazabicyclononanones, a new class of opioid-type Porzel, A. & Huneck, S. Acaranoic acid and acareno- Biochemistry and Molecular Biology 32, 199-204 Brandt, W. Development of a tertiary-structure lated leaves of transgenic tobacco plants expressing dwarf mutants. Plant Physiol. 125, 2053-2058 analgesics. Science and Culture 68, 11-18 (2002). ic acid: Confirmation of structure by modern NMR (2002). model of the C-terminal domain of DPPIV. In: Adv. yeast-derived invertase. Phytochemistry 56, 535- (2001). Methods. Bibliotheca Lichenologica 78, 365-368 Exp. Med. Biol., Vol 477, Cellular Peptidases in 541 (2001). Hui Xu, Zhao-Xin Wang, Schmidt, J., Heide, L. & (2001). Stano, J., Micieta, K., Neubert, K., Luckner, M. & Immune Functions and Diseases (2), (Langner, J. & Clemens, S., Schroeder, J. I. & Degenkolb, T. Shu-Ming Li. Genetic analysis of the biosynthesis of Adam, G. A simple method for the identification Ansorge, S., eds.) Kluwer Academic/Plenum Berger, S., Weichert, H., Porzel, A., Wasternack, C., Caenorhabditis elegans expresses a functional phy- the pyrrole and carbamoyl moieties of coumermy- Porzel, A., Trinh Phuong Lien, Schmidt, J., Drosihn, S., and assay of extracellular plant b-galactosidase. Publishers, Dordrecht, pp. 97-102 (2000). Kühn, H. & Feussner, I. Enzymatic and non-enzyma- tochelatin synthase. Eur. J. Biochem. 268, 3640 - cin A1 and novobiocin. Mol. Genet. Genomics 268, Wagner, C., Merzweiler, K., Tran Van Sung & Adam, Pharmazie 57, 176-177 (2002). tic lipid peroxidation in leaf development. Biochim. 3643 (2001). 387-396 (2002). G. Fissistigmatins A-D: Novel type natural products Bühling, F., Nägler, D., Fengler, A., Brandt, W., Welte, T. Biophys. Acta 1533, 266-276 (2001). with flavonoid-sesquiterpene hybrid structure from Tierens, K. F. M.-J., Thomma, B. P. H. J., Brouwer, M., & Ansorge, S. The growing family of mammalian Drosihn, S., Porzel, A. & Brandt, W. Determination Irmler, S., Schröder, G., St-Pierre, B., Crouch, N. P., Fissistigma bracteolatum. Tetrahedron 56, 865-872 Schmidt, J., Kistner, K., Porzel, A., Mauch-Mani, B., papain-like cysteine proteinases. In: Adv. Exp. Med. Berlich, M., Menge, S., Bruns, I., Schmidt, J., Schneider, of preferred conformations of brassinosteroids by Hotze, M., Schmidt, J., Strack, D., Matern, U. & (2000). Cammue, B. P. A. & Broekaert, W. F. Study of the Biol., Vol 477, Cellular Peptidases in Immune B. & Krauss, J. Coumarins give misleading absorban- means of NMR investigations and Boltzmann stati- Schröder, J. Indole alkaloid biosynthesis in Role of Antimicrobial Glucosinolate-Derived Functions and Diseases (2), (Langner, J. & Ansorge, ce with Ellman's reagent suggestive of thiol conju- stical analysis of simulated annaeling calculations. J. Catharanthus roseus: new enzyme activities and Samappito, S., Page, J., Schmidt, J. De-Eknamkul, W. & Isothio cyanates in Resistance of Arabidopsis thalia- S., eds.) Kluwer Academic/Plenum Publishers, gates. Analyst 127, 333-336 (2002). Mol. Model. 7, 34-42 (2001). identification of cytochrome P450 CYP72A1 as Kutchan, T. M. Molecular characterization of root- na to Microbial Pathogens. Plant Physiol. 125, 1688- Dordrecht, pp. 241-254 (2000). secologanin synthase. Plant J. 24, 797-804 (2000). specific chalcone synthases from Cassia alata. 1699 (2001). Blitzke, T., Baranovsky, A. & Schneider, B. Synthesis Eichelberger, U., Mansourova, M., Hennig, L., Planta 216, 64-71 (2002). Fengler, A. & Brandt, W. Development and validation and protein binding of (4-carboxybutyl)carbamoyl- Findeisen, M., Giesa, S., Müller, D. & Welzel, P. A cross Kamperdick, C., Nguyen Minh Phuong, Tran Van Tran Van Sung, Trinh Thi Thuy, Thach Thi Dan, Adam, of homology models of human cathepsins K, S, H, sustituted Taxoids. Helv. Chim. Acta 84, 1989-1995 metathesis-based synthesis of analogues of the 2- Sung & Adam, G. Guaiane dimers from Xylopia viel- Schliemann, W., Cai, Y., Degenkolb, T., Schmidt, J. & G. & Merzweiler, K. Isolation and structure of iso- and F. In: Adv. Exp. Med. Biol., Vol 477, Cellular (2001). O-alkyl glycerate part of the moenomycins. ana. Phytochemistry 56, 335-340 (2001). Corke, H. Betalains of Celosia argentea. corydin and corydalmin from the rhizome of Peptidases in Immune Functions and Diseases (2) Tetrahedron 57, 9737-9742 (2001). Phytochemistry 58, 159-165 (2001). Stephania rotunda. J. of Chemistry (Vietnamesisch) (Langner, J. & Ansorge, S., eds.) Kluwer Blitzke, T., Masaoud, M. & Schmidt, J. Constituents of Kobayashi, N., Schmidt, J., Nimtz, M., Wray, V. & 40, 35-40 (2002). Academic/Plenum Publishers, Dordrecht, pp. 255- Eulophia petersii. Fitoterapia 71, 590-591 (2000). Eichelberger, U., Neundorf, I., Hennig, L., Findeisen, Schliemann, W. Betalaines from Christmas cactus. Schmidt, J., Blitzke, T. & Masaoud, M. Structural inve- 260 (2000). M., Giesa, S., Müller, D. & Welzel, P. Synthesis of ana- Phytochemistry 54, 419-426 (2000). stigations of 5-methylchromone glycosides from Trinh Phuong Lien, Kamperdick, C, Schmidt, J., Tran Blitzke, T., Masaoud, M. & Schmidt, J. Constitutents logues of the 2-O-alkyl glycerate part of the moe- Aloe species by liquid chromatography / electro- Van Sung & Adam, G. Apotirucallane triterpenoids Mrestani-Klaus, C., Fengler, A., Faust, J., Brandt, W., of Aloe rubroviolacea. Fitoterapia 72, 78-79 (2001). nomycins. Tetrahedron 58, 545-559 (2002). Kobayashi, N., Schmidt, J., Wray, V. & Schliemann, W. spray tandem mass spectrometry. Eur. Mass. from Luvunga sarmentosa (Rutaceae). Wrenger, S., Reinhold, D., Ansorge, S. & Neubert, K. Metabolic formation and occurrence of dopamine- Spectrom. 7, 481-490 (2001). Phytochemistry 60, 747-754 (2002). N-terminal HIV-1 Tat nonapeptides as inhibitors of Blitzke, T., Porzel, A., Masaoud, M. & Schmidt, J. A Ettrich, R., Brandt, W., Kopecky Jr., V., Baumruk, V., derived betacyanins. Phytochemistry 56, 429-436 dipeptidyl peptidase IV. Conformational characteri- chlorinated amide and piperidine alkaloids from Hofbauerova, K. & Pavlicek, Z. Study of chaperone- (2001). Schmidt, J., Richter, K., Voigt, B. & Adam, G. Trinh Phuong Lien, Porzel, A. & Schmidt, J., Tran Van zation. In: Adv. Exp. Med. Biol., Vol. 477, Cellular Aloe sabaea. Phytochemistry 55, 979-982 (2000). like activity of human haptoglobin: Localisation of Metabolic transformation of the brassinosteroid Sung & Adam, G. Chalconoids from Fissistigma Peptidases in Immune Functions and Diseases (2), chaperone binding sites on the three-dimensional Kolbe, A., Fuchs, P., Porzel, A., Baumeister, U., Kolbe, 24-epicastasterone by the cockroach Periplaneta bracteolatum. Phytochemistry 53, 991-995 (2000). (Langner, J. & Ansorge, S., eds.) Kluwer Blitzke, T., Schmidt, J. & Masaoud, M. 7-O- structure of the H-chain deduced by knowledge- A. & Adam, G. Synthesis and crystal structure of americana. Z. Naturforsch. 55c, 233-239 (2000). Academic/Plenum Publishers, Dordrecht, pp. 125- 2 Methylaloeresin A - a new chromone glycoside based modeling. Biol. Chem., 383, 1667-1676 [26,27- H6] 24-epicathasterone. J. Chem. Soc., Trinh Phuong Lien, Tran Van Sung & Adam, G. 130 (2000). from Commiphora socotrana. Nat. Prod. Letters (2002). Perkin Trans. 1, 2022-2027 (2002). Schmidt, J. & Wessjohann, L. Studies in natural pro- Phytochemische Untersuchungen über 15, 27-33 (2001). ducts chemistry. Book reviews in Phytochemistry Inhaltsstoffe der vietnamesischen Heilpflanze Schmidt, J., Spengler, B., Voigt, B. & Adam, G. Franke, K., Kuhnt, C., Schmidt, J. & Munoz, O. 24- Kolbe, A., Kramell, R., Porzel, A., Schmidt, J., 61, 880 (2002). Tabernaemontana corymbosa (Nghien Cúu Thành Brassinosteroids - Structures, Analysis and Braga, A. L., Appelt, H. R., Schneider, P. H., Rodrigues, epi-castasterone and phytosterols from seeds of Schneider, G. & Adam, G. Synthesis of dexametha- Phàn Hóa Hoc Cay Lài Trau Tu Tán Synthesis. In: Evolution of Metabolic Pathways, O. E. D., Silveira, C. C. & Wessjohann, L. A. New C2- Maytenus boaria (Celastraceae). Rev. Latinoamer. sone conjugates of the phytohormones gibberellin Schneider, G., Fuchs, P. & Schmidt, J. Evidence for the Tabernaemontana Corymbosa). Zeitschrift für Recent Advances in Phytochemistry 34 (Romeo, J. 2 symmetric chiral disulfide-ligands derived from (R)- Quim. 27, 111-115 (2000). A3 and 24-epicastasterone. Collect. Czech. Chem. direct 2b- and 3b-hydroxylation of [ H2]GA20-13- Chemie Vietnam 39, 39-44 (2001). T., Ibrahim, R., Varin, L. & DeLuca, V., eds.) Pergamon, 2 cysteine. Tetrahedron 57, 3291-3295 (2001). Commun. 67, 103-114 (2002). O-[6'- H2]glucoside in seedlings of Phaseolus cocci- Amsterdam, pp. 385-407 (2000). Franke, K., Masaoud, M. & Schmidt, J. Cardanols neus L. Physiologia Plantarum 116, 144-147 (2002). Trinh Thi Thuy, Tran Van Sung & Adam, G. Limonoide Braga, A., Silva, S., Lütke, D., Drekener, R., Silveira, C., from Rhus thyrsiflora. Planta Medica 67, 477-479 Landtag, J., Baumert, A., Degenkolb, T., Schmidt, J., aus der vietnamesischen Heilpflanze Clausena Vasilets, L. A., Brandt, W., Postina, R., Fotis, H., Rocha, J. & Wessjohann, L. Chiral diselenide ligands (2001). Wray, V., Scheel, D., Strack, D. & Rosahl, S. Schneider, G., Koch, M., Fuchs, P. & Schmidt, J. excavata (Các Hop Chát Limonoit Tù Cay Hòng Bi Tatjanenko, L. V. & Gvozdev, A. R. Molecular mecha- for the asymmetric copper-catalyzed conjugate Accumulation of tyrosol glucoside in transgenic Identification of metabolically formed glucosyl con- Dai Clausena excavata). Zeitschrift für Chemie nisms of covalent regulation of the Na+/K+-ATPase addition of Grignard reagents to enones. Franke, K., Porzel, A., Masaoud, M., Adam, G. & potato plants expressing a parsley tyrosine decar- jugates of [17-D2]GA34. Phytochem. Anal. 11, 232- Vietnam 39, 27-33 (2001). by protein kinases. In: The Sodium Pump (Taniguchi, Tetrahedron Letters 43, 7329-7331 (2002). Schmidt, J. Furanocoumarins from Dorstenia gigas. boxylase. Phytochemistry 60, 683-689 (2002). 239 (2000). K. & Kaya, S. eds.) Elsevier, Amsterdam, pp. 507-572. Phytochemistry 56, 611-621 (2001). Vasilets, L. A., Brandt, W., Postina, R., Kirichenko, S.& Brandt, W. Struktur-Wirkungsbeziehungen von Lecaille F., Choe Y., Brandt W., Li, Z, Craik, C.S. & Schrekker, H., de Bolster, M., Orru, R. & Anders, A. (2000) Molecular mechanisms of PKC- Wessjohann, L. A. & Scheid, G. Synthetic Access to Opioiden. Pharmazie in unserer Zeit 31, 60-66 Franke, K., Porzel, A. & Schmidt, J. Flavone-coumarin Bromme, D. Selective inhibition of the collagenoly- Wessjohann, L. In Situ Formation of Allyl Ketones mediated inhibition of cation transport by the Epothilones - Natural Products with Extraordinary (2002). hybrids from Gnidia socotrana. Phytochemistry 61, tic activity of human cathepsin K by altering its S2 via Hiyama-Nozaki Reactions Followed by a Na+/K+-ATPase: sitedirected mutagenesis and Anticancer Activity. In: Organic Chemistry 873-878 (2002). subsite specificity. Biochemistry 41, 8447-8454 Chromium-Mediated Oppenauer Oxidation. J. Org. molecular modelling studies. Pflügers Arch. 439, Highlights IV (Schmalz, H.-G., ed.) Wiley-VCH, Brandt, W., Anders, A. & Vasilets, L. A. Predicted alte- (2002). Chem. 67, 1975-1981 (2002). R321 (2000). Weinheim, pp. 251-267 (2000). rations in tertiary structure of the N terminus of Galm, U., Schimana, J., Fiedler, H.-P., Schmidt, J., Shu- Na+/K+-ATPase alpha subunit caused by phosphory- Ming Li & Heide, L. Cloning and analysis of the Loc, Tran Van, Tran Van Sung, Kamperdick, C. & Schrick, K., Mayer, U., Horrichs, A., Kuhnt, C., Bellini, Voigt, B., Porzel, A., Adam, G., Golsch, D., Adam, W., Wrenger, S., Reinhold, D., Faust, J., Mrestani-Klaus, lation or acidic replacement of the PKC phospho- simocyclinone biosynthetic gene cluster of Adam, G. Synthesis of amino acid conjugates and C., Dangl, J., Schmidt, J. & Jürgens, G. FACKEL is a Wagner, C. & Merzweiler, K. Synthesis of 2,24-die- C., Brandt, W., Fengler, A., Neubert, K. & Ansorge, S. rylation site Ser-23. Cell Biochemistry and Streptomyces antibioticus Tü 6040. Arch Microbiol further derivatives of 3b-hydroxylup-20(29)ene- sterol C-14 reductase required for organized cell picastasterone and 3,24-diepicastasterone as Effects of nonapeptides derived from the N-termi- Biophysics 37, 83-95 (2002). 178, 102-114 (2002). 23,28-dioic acid. J. für praktische division and expansion in Arabidopsis embryogene- potential brassinosteroid metabolites of the cok- nal structure of human immunodeficiency virus-1 Chemie/Chemiker-Zeitung 342, 63-71 (2000). sis. Genes & Development 14, 1471-1484 (2000). kroach Periplaneta americana. Collect. Czech. (HIV-1) Tat on suppression of CD26-dependent T Bringmann, G., Schlauer, J., Rischer, H., Wohlfarth, M., Gao, W., Löser, R., Raschke, M., Dessoy, M., Fulhorst, Chem. Commun. 67, 91-102 (2002). cell growth. In: Adv. Exp. Med. Biol., Vol 477, Cellular Mühlbacher, J., Buske, A., Porzel, A., Schmidt, J. & M., Alpermann, H., Wessjohann, L. A. & Zenk, M. H. Lübken, T., Kraus, A. & Lorenz, W. Polyphenole in Schrick, K., Mayer, U., Martin, G., Bellini, C., Kuhnt, Peptidases in Immune Functions and Diseases (2), Adam, G. Revised structure of antidesmone, an (E)-4-Hydroxy-3-methylbut-2-enyl diphosphate: An Weinen aus Sachsen und Sachsen-Anhalt. C., Schmidt, J. & Jürgens, G. Interactions between Voigt, B., Whiting, P. & Dinan, L. The ecdysteroid ago- (Langner, J. & Ansorge, S. eds.) Kluwer unusual alkaloid from tropical Antidesma plants intermediate in the formation of terpenoids in Lebensmittelchemie 56, 103 (2002). sterol biosynthesis genes in embryonic develop- nist/antagonist and brassinosteroid-like activities of Academic/Plenum Publishers, Dordrecht, pp. 161- (Euphorbiaceae). Tetrahedron 56, 3691-3695 plant chromoplasts. Angew. Chem. Int. Ed. 41, 2604- ment of Arabidopsis. Plant J. 31, 61-73 (2002). synthetic brassinosteroid/ecdysteroid hybrid mole- 166 (2000). (2000). 2608 (2002). cules. Cell. Mol. Life Sci. 58, 1133-1140 (2001).
40 41 Publications, Books and Bookchapters, Publications in press, Research Group: Patents, Doctoral Theses, Diploma Theses Searching for Signals: Stress-Induced Changes in Arabidopsis Secondary Metabolite, Peptide and Protein Patterns (GABI) Heads: Stephan Clemens, Jürgen Schmidt, Ludger Wessjohann, Dierk Scheel
Publications in Press Nguyen Thi Hoang Anh, Tran Van Sung, Wessjohann, und Zanthoxylum avicennae (Rutaceae), National Eckermann, C., Schröder, G., Eckermann, St., Strack, L. & Adam, G. The iridoids and iridoid glucosid from Center for Natural Scientific and Technology Our project is aiming at contributing to the "post-genomic" ana-lysis of the Group members D., Schmidt, J., Schneider, B. & Schröder, J. Stilbene the Rehmannia glutinosa rhizome. J. of Chemistry (Vietnam), Institute of Chemistry and Martin- model organism Arabidopsis thaliana by establishing an extensive profiling of carboxylate biosynthesis: a new function in the (Vietnamesisch), in press (2002/2003). Luther-University of Halle-Wittenberg, Thomas Degenkolb family of chalcone synthase related proteins. 01/11/2001. proteins, peptides, and metabolites. These profiles are to be used for the (postdoctoral position since June 2000) Phytochemistry 62, 271-286 (2003). Samappito, S., Page, J., Schmidt, J., De-Eknamkul, W. detection and identification of early stress responses and novel signaling Claudia Horn & Kutchan, T. M. Aromatic and pyrone polyketides Wild, Harry: Neue Anwendungen der Chrom- (technician) Huneck, S., Lumbsch, H. Th., Porzel, A. & Schmidt, J. synthesized by a stilbene synthase from Rheum Reformatsky-Reaktion, Ludwig-Maximilians-Uni- molecules. Eventually, they will provide valuable tools for the analysis of Die Verteilung von Flechteninhaltsstoffen in tataricum. Phytochemistry 62, 313-323 (2003). versität München, Dept. of Organic Chemistry, Kerstin Körber Lecanora muralis und Lecidea inops und die 10/10/2000. various developmental and stress-induced changes as well as for the bioche- (technician) Abhängigkeit der Usninsäure-Konzentration vom Doctoral Theses mical phenotyping of mutants and the exploration of natural diversity. Substrat und von den Jahreszeiten bei Lecanora Buske, Alexander: Phytochemische Unter - Diploma Theses Edda von Röpenack-Lahaye muralis. Herzogia. suchungen der afrikanischen Euphorbiaceen Belting, Claudia: Terpene derived enediyes as Exemplary biotic and abiotic stresses under investigation are pathogen attack (postdoctoral position since May 2000) Antidesma membranaceum und Antidesma veno- potential anti-tumor drugs. Hochschule Enschede, and toxic metal exposure, respectively. Udo Roth Kolbe, A., Porzel, A., Schmidt, J. & Adam, G. A new sum. Martin-Luther-University of Halle- The Netherlands, 31/01/2002 (Sandwich). (postdoctoral position since May 2000) 2 synthesis of [26,28- H6]brassinolide and [26,28- Wittenberg, 19/10/2000. 2 H6]castasterone via unusual methyl migration. J. Bräuer, Lars: Modellierung der 4-Hydroxybenzoat Lab. Comp. Radiopharm. Frutos-Höner, Annabelle: Methodology studies on Oligoprenyltransferase und Charakterisierung in situ generated Fischer carbene complexes of potentiell aktiver Zentren. Martin-Luther- Collaborators Mrestani-Klaus, C., Brandt, W., Faust, J., Wrengler, S., group VI transition metals, on the Chromium- University of Halle-Wittenberg, 25/06/2002. The profiling of stress-induced metabolic on natural di ver sity, data sets for a num- Reinhold, D., Ansorge, S. & Neubert, K. New results Reformatsky reacion of nitrogen based com- changes in Arabidopsis plants grown ber of Ara bidopsis ecotypes are being ge - Thomas Altmann on the conformations of potent DP IV (CD26) inhi- pounds, and analytical studies of the Vogel collec- Spelbos, Vincent: Prenyltransferasen in Humulus under sterile conditions in a hydroponic nerated. University of Potsdam, Germany bitors bearing the N-terminal MWP structural tion, Sandwich between Ludwig-Maximilians- lupulus. University of Utrecht, The Netherlands, GABI-Arabidopsis-Verbund III motif. Int. Conf. "Dipeptidyl aminopeptidases: Basic Universität München, Dept. of Organic Chemistry November 2002. system has been established. A standardi- science and clinical applications", Berlin, 26.-29.09. and University of California in San Diego - UCSD Paul Schulze-Lefert, (USA), November 2000. Schültingkämper, Heike: The combinatorial diaste- zed extraction procedure for root and The profiling of proteins and peptides is Bernd Weisshaar Münzenberger, B., Hammer, E., Wray, V., Schauer, F., reoselective synthesis of highly functionalized leaf (secondary) metabolites is intro- based on two-dimensional gel electropho- Max Planck Institute of Plant Breeding Research, Schmidt, J. & Strack, D. Detoxification of ferulic acid Scheid, Günther: A New Synthesis of Epothilone tetrahydropyrans. Hochschule Enschede, The Cologne, Germany by ectomycorrhizal fungi. Mycorrhiza. Macrocycles, Free University of Amsterdam (NL), Netherlands, 18/01/2002 (Sandwich). duced. The methanolic extracts are analy- resis, MALDI-TOF-MS and nano-spray- Bioorganic Chemistry, 11/04/2002. zed by Cap-LC-ESI-Q-TOF-MS, hep- ESI-MS. Patterns of soluble leaf, Nguyen Thi Hoang Anh, Tran Van Sung, Wessjohann, Wesseling, Claudia: Synthesis of a building block for L & Adam, G. Some homoisoflavonoidal compounds Sinks, Udo Eckard: New Applications of Sulfoxides the natural product cis-gigantrionenine. tane extracts for the more hy - root or seed proteins are from Ophiopogon japonicus Ker - Gawler. J. of and Synthesis of Asymmetric Phenylselenide Hochschule Enschede, The Netherlands, dro phobic com pounds are resolved in large-format Chemistry (Vietnamesisch), in press (2002/2003). Reagents, Sandwich between Free University of 31/01/2002 (Sandwich). Amsterdam (NL), Bio-organic Chemistry, and ana lyzed by GC-MS. Cap-LC- two-dimensional gels. Gel Nguyen Thi Hoang Anh, Tran Van Sung, Wessjohann, Ludwig-Maximilians-Universität München, Dept. of In the report period, another nine students recei- ESI-Q-TOF-MS represents a images are carefully ana- L. & Adam, G. Some hydroxycinamic acid esters of Organic Chemistry, 15/05/2001. ved their diploma-degree (dutch: Drs.) at the Free phenylethyl alcohol glycosides from Rehmannia glu- University of Amsterdam, Bio-organic Chemistry, new profiling approach that is lyzed. Interesting protein tinosa Libosh. J. of Chemistry (Vietnamesisch), in Trinh Thi Thuy: Phytochemische Untersuchungen under the supervision of Prof. Wessjohann.< to complement the more esta- spots showing stress-related press (2002/2003). der vietnameschen Heilpflanzen Clausena excavata blished GC-MS techniques. Because chan ges in abundance are picked, di - very few tools are available for data gested and subjected to MALDI-TOF deconvolution and data extraction we mass spectrometry for identification developed respective procedures for the based on peptide mass fingerprints. Image automatic data analysis. Several samples analysis, which represents the bottleneck can now be processed per day. In leaf of searches for changes within the pro- extracts about 1200 mass signals are teome, has been optimized by adopting resolved and detected, in root signals new ima ging software. A number of Ara - about 1000 mass signals. Mass data can be bidopsis mutants have been analyzed un - directly compared to an Arabidopsis lite- der different stress conditions. For some of rature database. The exceptional mass the identified proteins that show stress- accuracy of ESI-Q-TOF-MS together with re lated changes functional characterization its tandem MS option allows tentative has been initiated by isolating Arabidopsis identification or classification of intere- insertion lines for the respective genes. sting compounds. An extensive evaluation of the whole Cap-LC-ESI-Q-TOF-MS- The intercellular washing fluid of Ara - based profiling approach is now complete. bidopsis leaves is analyzed for peptides Changes in response to the stress caused and metabolites by nanospray-ESI-Q- by exposure to elevated heavy metal TOF-MS. In principle, the detection of levels have been analyzed. In the course of molecules in this compartment is possi- collaborations with other groups, several ble. Progress, however, has so far been figures: Reproducible plant growth of Arabidopsis Arabidopsis mutants with defects in, for slow due to limitations in MS capacity instance, signal transduction or de fense, as the same machine is used for cap- are being profiled. Similarly, in order to be LC-coupled metabolite profiling and able to use metabolite profiling for studies nanospray-MS. <
42 43 Department: Stress and Developmental Biology Head: Prof. Dierk Scheel Secretary: Ruth Laue
lant development, although gene- Ptically determined, is largely modulated by biotic and abiotic envi- ronmental factors. In this way, deve- lopmental programs are adapted to specific lo cal con ditions and protecti- v e as well as d e fe n s e reac tions are initia- ted du ring stress situations - an advantageous situation for sedentary living plants.
The basis for those processes is the ability of plants to perceive environ- mental factors and ini-tiate signal transduction networks that modify gene expression patterns. The inve- stigation of the molecular mecha- nisms underlying this course of events is the main topic of the department of "Stress and Deve lop mental Bio logy".
Plant patho gens play a major role in bio- tic stress. The work of several rese- arch groups of the department focuses on the ana-lysis of recogni- tion, signal transduc- tion and gene activation processes in plant-pathogen interactions. The work on abio- tic environmental factors centers around metal homeostasis in plants, using hyperaccumulating model orga- nisms. <
44 45 Research Group: Signal Perception in Plant-Pathogen Interactions Head: Thorsten Nürnberger
Group members Innate immunity is well described for animals and is also suggested to be plants. (a) (b) Frédéric Brunner important for plants. In vertebrates and insects, microbial pathogen sensing In addition, all Phytophthora species (postdoctoral position) relies on the recognition of pathogen-specific structures, which are not found tested possess a 24-kDa protein (NPP1) Jutta Elster in hosts and which are indispen-sable for the lifestyle of the microorganism. that triggers defense responses in pars- (technician) ley very similar as does Pep-13. NPP1- Stephan Engelhardt Receptor-mediated signal perception by the host gives rise to the activation mediated activation of pathogen defense (student since November 2002) of specific immune responses, such as the synthesis of antimicrobial com- in parsley does not employ the Pep-13 Guido Fellbrich (PhD student until July 2002) pounds. We investigate whether pathogen recognition by animals and plants receptor. However, early-induced cellular Yvonne Gäbler share similar characteristics. Our data suggest that the evo lution of pathogen responses implicated in elicitor signal (student until 2001, afterwards PhD student) perception systems in plants is likely to be si mi lar to that described for transmission (increased levels of cyto- Claudia Horn animals. Microbial surfaces constitute com plex patterns for the activation of plasmic calcium, production of reactive (technician) oxygen species, MAP kinase activation) Birgit Kemmerling plant pathogen defense. Recognition of microbial pattern by plants appears to were stimulated by either elicitor, sugge- stably asso ciate with liposomes and syn- C-terminal fraction of the protein in (postdoctoral position since April 2002) result in more sensitive perception of pathogens and synergistically enhanced sting the existence of converging signa- thetic bilayer membranes. Under sym- both plants. Thus, pore-forming activity Justin Lee (postdoctoral position) plant defense. Phytopathogenic bacteria of the genus Pseudomonas produce ling pathways in parsley. Infiltration of metric ionic conditions, addition of 2 nM of HrpZPsph does not determine the acti- Annette Romanski and secrete the effector protein HrpZ during (attempted) infection of plants. NPP1 into leaves of Arabidopsis thaliana purified recombinant HrpZPsph to the cis- vation of plant defense, but may reflect (PhD student until July 2001) HrpZ was shown to insert into lipid bilayer mem branes and to form cation- resulted in transcript accumulation of compartment of planar lipid bilayers pro - the role of the protein during (attemp- Christel Rülke pathogenesis-related (PR) genes, produc- voked an ion current with a large uni tary ted) bacterial infection of plants. < (technician) conducting channels. This ion channel-forming activity, however, appears not tion of reactive oxygen species and ethy- conductivity of 207 pS. HrpZPsph-related to be the molecular basis for the activation of defense responses in plants tre- lene, callose apposition, and hypersensi- proteins from P. s. pv. tomato or syringae ated with HrpZ. Collaborators tive-like cell death. NPP1-mediated in - triggered ion currents similar to those duction of the PR1 gene is salicylic acid- stimulated by HrpZ . The HrpZ - Guy Cornelis Psph Psph (c) University of Brussels, Belgium dependent, and, unlike the P. syringae pv. mediated ion-conducting pore was per- Georg Felix tomato DC3000(avrRpm1)-induced PR1 meable for cations but did not mediate Friedrich Miescher Institute, Basel, Switzerland A calcium-dependent transglutaminase cultivar-specific defense responses in gene expression, required both func - fluxes of Cl¯. Such pore-forming activity Jane Glazebrook, Tong Zhu (TGase) present in the cell wall of as parsley and potato.An evolutionarily high- tional NDR1 and PAD4. Importantly, may allow nutrient release and/or deli- Torrey Mesa Research Institute (Syngenta), many as ten species of the genus ly conserved peptide fragment of this Ara bi dopsis plants infiltrated with NPP1 very of virulence factors during bacterial San Diego, USA Phytophthora serves as recognition protein (Pep-13) was identified within a constitute an experimental system that is coloni zation of host plants. In addition, Heribert Hirt University of Vienna, Austria determinant for the activation of non- surface-exposed loop structure of the amenable to forward genetic approaches HrpZ has been shown to trigger a com- Sakari Kauppinen, protein. Pep-13 was aiming at the dissection of signaling path- plex defense response in parsley and Grete Rasmussen shown to be neces sary ways implicated in the activation of non- tobacco. Ligand / receptor interaction NOVO NORDISK A/S, Bagsvaerd, Denmark and sufficient for re - cultivar-specific plant defense. studies revealed the presence of a high- Harald Keller ceptor-mediated activa- affinity binding site for HrpZPsph in plasma Institut National de la Recherche Agronomique (INRA), Antibes, France tion of de fense responses The HrpZ gene product from the bean membranes of both plants. Series of Birgit Klüsener, Elmar Weiler in both plants. Mutations halo-blight pathogen, Pseudomonas truncated HrpZPsph proteins were analy- NPP1 induces a complex defense response in University of Bochum, Germany within the Pep-13 motif of syringae pv. phaseolicola (HrpZPsph), is zed with respect to their abilities to Arabidopsis thaliana Col-0. Infiltrations were per- John Mansfield the P. sojae TGase, which secreted in an Hrp-dependent manner induce plant defense as well as to form formed with 2.5 µM of each recombinant NPP1, a Imperial College at Wye, University of London, UK mutant derivative of NPP1 with reduced activity, reduced or abolished the by this bacterium, and exported by the ion-conducting pores in liposomes. The or Glutathione-S-Transferase as control. Necrotic Nicholas Panopoulos lesion formation 48 h upon elicitation (a), produc- University of Crete, Greece elicitor activity of the in - type III se cretion system when expres- pore-forming activity of HrpZPsph was tion of reactive oxygen species 3 h upon elicita- tact protein, similarly af - sed in the mammalian pathogen Yersinia found to require the intact protein, while tion (b), and callose apposition 24 h upon elicita- Steffen Panzner tion (c). Novosom AG, Halle, Germany fected its enzyme activity. enterocolitica. HrpZPsph was found to defense responses were stimulated by a Martin Romantschuk Apparently, during evolu- University of Helsinki, Finland tion plants have acquired Dietmar Stahl receptors for the recog - Kleinwanzlebener Saatzucht AG, Einbeck, Germany nition of stable and func- Zhongmin Wei tionally indispensable sur- EDEN Bioscience, Bothell, USA NPP1-induced cell death in parsley Viability of parsley protoplasts treated with 20 nM NPP1 or water face epitopes of micro bial (control) was determined 24 h upon elicitation (upper panel). Viability patho gens, suggesting that of parsley protoplasts (5x105/ml) was determined by double-staining with 50 µg/ml fluorescein diacetate and 10 µg/ml propidium iodide 24 h such perception mo dules after treatment (Jabs et al., 1997). may form the molecular NPP1 (2.5 µM) or water (control) infiltrated into parsley leaves for 48 h basis of durable pathogen (lower panel). resistance in non-host
46 47 Research Group: Cellular Signaling Head: Dierk Scheel
Group members Parsley is not a host plant for the soybean pathogen, Phytophthora sojae, but if While the activation of MAPKs and PR1, ratory burst oxidase, these proteins are precursor 12-oxo-phytodienoic acid. Si - germinating zoospores of this oomycete try to invade the plant, it responds PR2 and WRKY transcription factor N-terminally extended by a region har- multaneous treatment of the cells with Reetta Ahlfors 2+ (guest scientist since July 2002, PhD student) with a multifaceted defense response that terminates the infection process.The genes was found to be independent of boring two Ca -binding EF hands. One lipoxygenase inhibitors completely blok- Barbara Degner oligopeptide elicitor Pep-13, originating from a hyphal cell wall transglutaminase the oxydative burst, superoxide anion of the NADPH oxidase transcripts accu- ked the accumulation of both oxylipins, (technician) of P. sojae, is one of the pathogen-associated molecular patterns (PAMPs) radicals, the primary reactive oxygen mulates rapidly and transiently upon eli- but did not affect Pep-13-mediated phy- Magdalena Krzymowska species formed during the Pep-13-stimu- citation. In addition, transcripts encoding toalexin synthesis, suggesting that jasmo- (postdoctoral position until June 2002) recognized by the plant cell via a plasma membrane-localized receptor (see pre- ceding report). Upon binding of Pep-13, this receptor initiates a cellular signal lated oxidative burst, are necessary and enzymes with and without EF hands nate and/or 12-oxo-phytodienoic acid Violetta Macioszek sufficient for phytoalexin production and were found to be generated by alterna- re present the starting point of yet anot- (postdoctoral position since September 2002) transduction cascade that causes dramatic alterations of the gene expression activation of those genes encoding their tive splicing. Heterologous expression of her signal transduction branch. Salicylic Anja Nickstadt pattern, primarily resulting from activation of defense-related genes. The cellu- (PhD student until May 2002) biosynthetic and additional phenylpropa- both type of proteins in yeast resulted in acid, a plant defense signaling compound lar signaling elements linking the Pep-13 receptor to specific activation of Jason Rudd noid pathway enzymes. The formation of production of active NADPH oxidases involved in signaling pathways that initia- (postdoctoral position since April 2000) defense-related genes include plasma membrane-located ion channels, protein superoxide anion radicals is catalyzed by embedded in microsomal membranes. te programmed cell death, does not Rita Schlichting kinases, an NADPH oxidase and jasmonate. Together with additional unknown NADPH oxidases, which are structurally Only the larger protein with the EF accumulate in Pep-13-treated parsley (PhD student since July 2002) components, these elements form a modular signaling network tightly regula- similar to the catalytic subunit of the hands required Ca2+ for activity. cells. Interestingly, parsley cells and leaves Heidi Zinecker mammalian respiratory burst oxidase. do not undergo programmed cell death (PhD student until December 2000) ting the temporal and spatial activation of defense reactions. Two NADPH oxidase-encoding genes The oxidative burst is necessary but not in response to Pep-13 treatment. < were isolated from parsley. In compari- sufficient for Pep-13-stimulated produc- Collaborators son to the catalytic subunit of the respi- tion of the oxylipins, jasmonate and its Thomas Boller Pep-13 treatment of suspension-cultured been isolated from parsley, designated 2+ Friedrich Miescher Institute, Basel, Switzerland parsley cells rapidly stimulates Ca influx PcMPK3a, 3b, 4 and 6 according to their Jeff Dangl resulting in a characteristic sustained sequence similarities to MAPK-encoding University of North Carolina, Chapel Hill, USA increase in cytosolic Ca2+ levels, which is genes of Arabidopsis thaliana. Upon elici- Jerome Giraudat essential for all the other known elicitor tation PcMPK3a, 3b, 6 and a fourth so far Institut des Sciences du Végétal, CNRS, Gif-sûr-Yvette, France responses. At least four mitogen-activa- unknown MAPK were found to be activa- Heribert Hirt ted protein kinase (MAPK) cascades are ted by phosphorylation of the conserved University of Vienna, Austria activated downstream of this Ca2+ tran- TEY motif and translocated to the Jonathan Jones sient. Four MAPK-encoding genes have nucleus, whereas PcMPK4 was not affec- The Sainsbury Laboratory, Norwich, UK ted. Transient co-expres- Chris Lamb sion of dominant inactive John Innes Centre, Norwich, UK versions of PcMPK3a, 4 John Mundy University of Copenhagen, Denmark and 6 with reporter gene Teun Munnik fusions of the PR1 (patho- University of Amsterdam, The Netherlands genesis-related) and PR2 Karsten Niehaus pro moters has demonstra- University of Bielefeld, Germany ted that elicitor activation Jane Parker, Imre Somssich of these genes is regulated Max Planck Institute for Plant Breeding Research, Cologne, Germany by PcMPK3a (and possibly Elicitor treatment induces nuclear Jose Sanchez-Serrano also 3b) and/or PcMPK6, translocation of MPK3 Autonomous University, Madrid, Spain but not by PcMPK4. Cultured parsley cells were treated with Pep13 (100 nM; C, D) or H2O MAPKs are themselves ac - (A, B) and fixed in 4% paraformaldehy- de 15 min. after initiation of treat- ti vated through phospho- ment. Cells were embedded in paraf- rylation by MAPK kinases. fin, cut into 6 µm sections and stained with PcMPK3 antiserum (A, C). Goat Two MAPK kinase-encoding anti-rabbit secondary antibody conju- genes, PcMEK1 and 2, have gated with Alexa 488 was used to visualize the primary antiserum bound been isolated from parsley. to MPK3; nuclei were also counter- Only PcMEK2 was found stained with DAPI (B, D). After treat- ment with Pep13 most nuclei were to be activated in Pep-13- stained by PcMPK3 antiserum, where- treated cells and was then as no or little nuclear staining was able to phosphorylate detectable in control cells. PcMPK3a, 3b and 6, not Pep-13-initiated signal transduction processes in parsley. however PcMPK4.
48 49 Research Group: Induced Pathogen Defense Heads: Sabine Rosahl & Dierk Scheel
Group members To elucidate defense mechanisms against the oomycte Phytoph- dies against jasmonic and 12-oxo-phyto- penetration is only observed in rare dienoic acid. cases. The plant cell reacts with the Carola Geiler t hora infestans, the causal agent of late blight disease of potato, we are study- (student until August 2001) ing the interaction of P. infestans with its host plant potato and with the non- Jasmonate-dependent expression of the deposition of callose, accumulation of Cornelia Göbel host plant Arabidopsis thaliana. For potato, analysis of the recognition of the proteinase-inhibitor-II-genes is reduced autofluorescent material and localized (PhD student until August 2001) pathogen, signal transduction and characterization of the pathogen defense in the transgenic plants indicating that hypersensitive cell death. The Arabi - Anja Grohnert are our major interests. the levels of physiologically active jasmo- dopsis mutant pen2 (collaboration with (student until February 2001) nic acid are reduced due to binding by Volker Lipka and Paul Schulze-Lefert, Vincentius A. Halim the antibodies. The effect on defense MPI Cologne), identified as allowing (PhD student since October 2002) The Phytophthora sojae-derived oligo- died. gene expression and on the response to enhanced penetration of Blumeria Astrid Hunger (PhD student until June 2002) peptide elicitor Pep-13, originally identi- Oxylipins play an important role in the pathogen infection is presently being gramnis f. sp. hordei, reacts similarly to Martina Kausch fied as an inducer of plant defense in plant's reaction to pathogen attack. In analyzed. P. in-festans infection with higher pene- (student until February 2002) parsley and shown to act as a pathogen- potato, 9-lipoxygenase-derived oxylipins tration frequencies and increased cell Jörn Landtag associated molecular pattern (PAMP) in accumulate in response to Pep-13 and Since P. infestans is not able to success- death. Although the first layer of de- (student until June 2001, PhD student since September 2001) evoking innate immune responses, also elicitor treatment as well as after patho- fully infect A. thaliana, the analysis of fense in nonhost resistance appears to Claudia Reh triggers defense responses in potato. In gen infection. To analyze the role of this nonhost pathogen interaction be affected in the mutant, pen2 is still (student until February 2000) cultured potato cells, Pep-13 treatment 9-lip oxy genase-derived oxylipins, trans- should elucidate mechanisms of defense able to contain the pathogen. To iden- Grit Rothe results in the formation of hydrogen genic potato plants expressing RNA against the infectious agent of late blight tify further components involved in (postdoctoral position since March 2002) peroxide, alkalinization of the culture interference constructs, targeted at the di-sease. Microscopic analyses revealed nonhost resistance, pen2 seeds were Angelika Weinel medium, accumulation of 9-lipoxygena- pathogen-induced 9-lipoxygenase of po - that P. in-festans spores germinate on mutagenized and are presently being (technician) se-derived oxylipins and activation of tato, were generated and are being ana- Arabidopsis leaves and attempt to screened for alterations in their Lore Westphal (postdoctoral position since April 2002) defense genes. Similarly, accumulation of lyzed for alterations in their response to pene trate cells. However, successful response to P. infestans infection. < transcripts encoding enzymes of the pathogen infection. Whether oxylipins phenylpro-panoid pathway, lipoxygenases from solanaceous plants like potato can Collaborators and pa tho genesis-related proteins also be effective against pathogens in Udo Conrad, Patrick Schweizer occurs in po tato leaves in response to other plants is being tested by transfer- Institute of Plant Genetics and Crop Plant Research, Pep-13 infiltration. Derivatives of Pep-13 ring the respective genes from potato Gatersleben, Germany show similar elicitor activity in parsley into A. thaliana. Ivo Feussner University of Göttingen, Germany and potato, suggesting a receptor-media- Markus Frank ted induction of defense response in The 13-lipoxygenase products jasmonic BASF Plant Science, Ludwigshafen, Germany potato analogous to that observed in acid and its precursor 12-oxo-phyto- Bettina Hause, Dieter Strack, parsley. Interestingly, unlike in parsley, dienoic acid accumulate in potato in Claus Wasternack infiltration of Pep-13 into leaves leads to response to infiltration of the phytopa- Institute of Plant Biochemistry, Halle, Germany rapid cell death in potato. Using transge- thogenic bacteria Pseudomonas syringae Volker Lipka, Jane Parker, Paul Schulze-Lefert nic plants with modulated levels of jas- pv. maculicola. This nonhost pathogen Max Planck Institute of Plant Breeding Research, monic and salicylic acid, the dependence interaction leads to local and systemic Cologne, Germany of Pep-13-induced defense reactions on de-fense gene expression and to increa- Mats Hamberg these signaling compounds is being stu- sed resistance against subsequent patho- Karolinska Institute, Stockholm, Sweden Immunolocalization of single chain antibodies gen attacks. 12-oxo-phytodienoic acid, directed against 12-oxo-phytodienoic acid in chlo- roplasts of transgenic potato plants. In contrast to but not jasmonic acid accumulates also untransformed control plants (upper panel), trans- systemically. To analyze the role of genic plants express single chain antibodies in chlo- roplasts as indicated by the green fluorescence these (middle and lower panel; B. Hause). 13-lipoxygenase products for defense responses, transgenic plants were gene- rated which express single chain antibo-
Growth of P. infestans in potato. The oomycete P. infestans, the cau- sal agent of late blight disesase of potato, spreads intercellularly in Arabidopsis thaliana plants are grown under con- susceptible potato leaves (left trolled conditions in a phytochamber. They are sub- panel) and is able to sporulate sequently screened for alterations in their respon- (right panel). Infected cells react se to infection with P. infestans, the causal agent of with cell death as indicated by the late blight disease of potato. Thousands of plants white autofluorescence (middle must be screened to obtain one mutant. panel).
50 51 Research Group: Metal Homeostasis Heads: Dieter Neumann & Stephan Clemens
Group members Plants - like all other organisms - are able to tightly regulate the intracellular na. A number of specifically metal-regula- insights into mechanisms of Zn homeosta- concentration and the distribution of essential heavy metals such as zinc and ted putative signal transduction compo- sis, Cd toxi city, and intracellular metal Clarice de Figuereido (PhD student) copper. Also, the cytosolic concentrations of non-essential toxic heavy metals nents have been identified both in A. hal- distribution. Marina Häußler (e.g. cadmium, lead) have to be minimized. Some plant species (so-called metal- leri and A. thaliana. Five of them are stu- Plant metal tolerance is an element-speci- (technician) lophytes) can tolerate otherwise toxic concentrations and grow on heavy died in detail. A. thaliana knock-out lines fic process. For cell cultures of Silene Emiko Harada have been obtained. Their metal responses jenissiensis it was shown that Zn and Cu (postdoctoral position since April 2002) metal contaminated soil. Main objective of the group is to elucidate the mecha- nisms underlying plant metal homeostasis and metal hyperaccumulation. We and possible metal-related phenotypes are de toxified by distinct mechanisms. Zn Elke Hillert are studied using a variety of techniques tolerance is mediated by two different Si- (technician) are using analytical electron microscopy and a range of biochemical and mole- including microarrays. Since recently it dependent processes. Exposure to Zn Sylvia Krüger cular techniques. Plants under investigation are Arabidopsis thaliana, its close (technician) was shown that Zn and Cd hyperaccumu- results in elevated Si content of cells. Zn relative Arabidopsis halleri, and other metallophytes (Silene vulgaris, Minuartia Thomas Maier lation by A. halleri is a con stitutive pheno- and Si containing precipitates are detec- (postdoctoral position until May 2002) verna and Armeria maritima). In addition, we are working with menon found also on soil with normal table in the cytosol and the mitochondria. Claudia Simm Schizosaccharomyces pombe as a cellular model for metal homeostasis. metal content, the molecular analysis was They were identified as incompletely sub- (PhD student since October 2000) extended to constitutive differences bet- stituted Zn silicate. Such silicates are un - Pierre Tennstedt ween the two Ara bi dopsis species. Gene stable and decompose to SiO , detec- (PhD student since August 2002) 2 expression profiles were obtained for table in the cytosol as an electron trans- Christoph Vess (PhD student) The formation of phytochelatins (PCs) is a tha liana was demonstrated. PCS genes are roots of hydroponically grown plants by parent structure and identified by EEL Susan Wassersleben principle response of plants, many fungi constitutively expressed. PC synthesis is using Affymetrix GeneChips. They revea- spectra. Zn silicate is hypothesized to (PhD student since July 2000) and algae to toxic metal exposure. We directly activated by the binding of a heavy led a number of about 20 genes which are func tion as a temporary storage form of Michael Weber showed that invertebrates such as metal ion or the corresponding glutathione significantly more active in A. halleri. Zn, which prevents toxic effects within Zn-silicate in mitochondria and cytoplasm of (PhD student since March 2001) chelate to the enzy- Among them are genes encoding several the cytosol. A second, unusual mechanism Silene jenessiensis (ESI). Uta zur Nieden me. In order to fur - known metal homeostasis factors such as may contribute to the Zn tolerance of (research scientist) ther elucidate this metal transporters and enzymes involved some plants. Apparently, a large fraction ac tivation process and in metal chelator synthesis. These genes of Zn is directly taken up into the va cuole Collaborators to establish a tech - represent prime candidates for determi- as Zn silicate without membrane passage. Udo Conrad, Renate Manteuffel nique for the cha- nants of Zn/Cd hyperaccumulation and Transport occurs in vesicles formed by Institute of Plant Genetics and Crop Plant Research, racterization of me - are therefore studied in detail. plasma membrane and tonoplast. Cu ex - Gatersleben, Germany tal-binding sites we posure, on the other hand, does not result Klaus Kloppstech used "peptide scans", Several putative metal tolerance factors in ultrastructu- University of Hannover, Germany i.e. spotted peptide are investigated in S. pombe, the model ral changes. Ute Krämer Max Planck Institute of Molecular Plant Physiology, libraries re presenting for PC-forming cells. The analysis of a Highest Cu con - Golm, Germany PCS proteins. Cd2+- transporter belonging to the Cation centrations are Gerhard Küllertz bin ding sites could be Diffusion Facilitators and of the only S. found in the Max Planck Research Unit for Enzymology of Protein Folding, Halle, Germany lo ca lized and functio- pombe me tallothionein has led to new mito chon dria Olaf Lichtenberger nally characterized by and there is no Institute of Plant Biochemistry, Halle, Germany site-directed mu ta - co-localization Enrico Martinoia genesis. with Si. < University of Neuchatel, Switzerland The phytochelatin pathway and possible other mechanisms of Cd2+ detoxifica- tion. PC synthesis from GSH is activated by several metal and metalloid ions. Dietrich Nies 2+ 2+ 2+ 2+ University of Halle, Germany Cd ions enter the cell via Fe , Zn or Ca transporters. Upstream of GSH The molecular ana - biosynthesis are sulphate assimilation and cysteine biosynthesis. In S. pombe, Uwe Schmidt PC-Cd complexes (LMW) are transported into the vacuole by the ABC-type lysis of plant metal Federal Research Centre for Forestry and Forest transporter Hmt1. In plant cells , this transport is hypothesized to be media- responses and deter- Products, Hamburg, Germany ted by a protein of the same family. Inside the vacuole HMW complexes are formed by addition of sulphide, which apparently is derived from cysteine. minants of metal hy - Julian Schroeder Other mechanisms of Cd2+ detoxification discussed for plants and other orga- per accumulation is University of California at San Diego, La Jolla, USA nisms are vacuolar sequestration dependent on either CDF proteins or 2+ + being pursued in the Wilhelm Schwieger Cd /H antiporters, binding to metallothioneins or efflux mediated by CPx- University of Erlangen, Germany type ATPases. model sys tems Arabi dopsis thaliana Caenorhabditis elegans also express func- and Arabi dopsis halleri. Ex pression profi- tional PC synthases (PCS) by expressing ling in A. halleri by cDNA-AFLP has been the respective protein in a PCS-deficient con tinued. The focus is on metal sensing Growth of two Arabidopsis species on normal (-Cd) and Cadmium- contaminated (+Cd) soil. Arabidopsis thaliana (left) growth is affec- S. pombe strain. In a similar way, the exi- and metal signal transduction since virtual- ted, whereas Arabidopsis halleri (right) is able to tolerate high stence of a second PCS in Arabidopsis ly noth ing is known about these phenome- Cadmium concentrations.
52 53 Publications, Books and Bookchapters, Publications in press, Patents, Doctoral Theses, Diploma Theses
Publications induction and modulation of plant defense respon- ses: versatile plant signalling components necessary Varet, A., Parker, J., Tornero, P., Nass, N., Nürnberger, A., eds.) Kluwer Academic Publishers Dordrecht. Biochemistry/Biotechnology, 21/11/2001. Abel, S., Nürnberger, T., Ahnert, V., Krauss, G.-J. & ses by bacterial lipopolysaccharides. Annu. Rev. for pathogen defence. Trends Plant Sci. 7, 97 T., Dangl, J. L., Scheel, D., Lee, J. NHL25 and NHL3, Neumann, D. Silicon in plants. In: Progress in Glund, K. Induction of an extracellular cyclic nucle- Phytopathol. 38, 241-261 (2000). (2002). two NDR1/HIN1-like genes in Arabidopsis thaliana Molecular and Subcellular Biology. Silicon otide phosphodiesterase as an accessory ribonu- with potential role(s) in plant defense. Mol. Plant 15 Biomineralization. Springer-Verlag, Wien-New York. Zinecker, Heidi: Reaktive Sauerstoffspezies in der cleolytic activity during phosphate starvation of cul- Fellbrich, G., Blume, B., Brunner, F., Hirt, H., Kroj, T., Li, J., Nass, N., Kusaba, M., Dodds, P., Treloar, N., Microbe Interact. , 608-616 (2002). pflanzlichen Pathogenabwehr - Isolierung und tured tomato cells. Plant Physiol. 122, 543-552 Ligterink, W., Romanski, A. & Nürnberger, T. Clarke, A. E. & Newbigin, E. J. A genetic map of the Nürnberger, T. Elicitor-mediated signal transduction Charakterisierung von Genen aus Petroselinum cri- (2000). Phytophthora parasitica elicitor-induced reactions Nicotiana alata S-locus that includes three pollen- Veit, S., Wörle, J. M., Nürnberger, T., Koch, W. & Seitz, in the activation of plant pathogen defense. In: Plant spum L., die für putative NADPH-Oxidasen kodie- in cells of Petroselinum crispum. Plant Cell Physiol. expressed genes. Theor. Appl. Genet. 100, 956-964 H. U. A novel protein elicitor (PaNie) from Pythium Hormone Research, Vol. 13, (Bisseling, T. & Schell, J., ren. University of Halle-Wittenberg, Department of Berger, S., Weichert, H., Porzel, A., Wasternack, C., 41, 692-701 (2000). (2000). aphanidermatum induces dual defense responses in eds.) Springer-Verlag, Wien-New York. Biochemistry/Biotechnology, 09/01/2001. Kühn, H. & Feussner, I. Enzymatic and non-enzyma- carrot and Arabidopsis. Plant Physiol. 127, 832-841 tic lipid peroxidation in leaf development. Biochim. Fellbrich, G., Romanski, A., Varet, A., Blume, B., Lubaretz, O. & zur Nieden, U. Accumulation of plant (2001). Patents Diploma Theses Biophys. Acta 1533, 266-276 (2001). Brunner, F., Engelhardt, S., Felix, G., Kemmerling, B., small heat-stress proteins in storage organs. Planta Books and Book chapters Feussner, I., Hornung, E. & Rosahl, S. 11-Ara - Gäbler, Yvonne: Anlage von cDNA-Microarrays der Krzymowska, M. & Nürnberger, T. NPP1, a 215, 220-228 (2002). chidonat-Lipoxygenase-Mutante. German patent Petersilie (Petroselinum crispum). University of Berger, S. Jasmonate-related mutants of Arabidopsis Phytophthora-associated trigger of plant defense in Bruns, I., Sutter, K., Neumann, D. & Krauss, G.-J. 19931819.0 (2001). Halle-Wittenberg, Department of Biochemis try/ as tools for studying stress signaling. Planta 214, parsley and Arabidopsis. Plant J. 32, 375-390 (2002). Luderer, R., Rivas, S., Nürnberger, T., Mattei, B., Van Glutathione accumulation - a specific response of Bio technology, 05/09/2001. 497-504 (2002). den Hooven, H. W., Van der Hoorn, R. A. L., Romeis, mosses to heavy metal stress. In: Sulfur Nutrition Scheel, D., Rosahl, S., Strack, D. & Schmidt, A. Trans - Göbel, C., Feussner, I., Schmidt, A., Scheel, D., T., Wehrfritz, J.-M., Blume, B., Nennstiel, D., and Sulfur Assimilation in Higher Plants (Brunold, gene Pflanzen mit erhöhter Resistenz gegen den Geiler, Carola: Induktion von Abwehrreaktionen in Berger, S., Mitchell-Olds, T. & Stotz, H. U. Local and Sanchez-Serrano, J., Hamberg, M. & Rosahl, S. Zuidema, D., Vervoort, J., De Lorenzo, G., Jones, J. D. C., ed.) Haupt, Bern, pp. 389-391 (2000). Befall durch Phytopathogene. German patent Solanum tuberosum L. durch den Oligopeptid- differential control of vegetative storage protein Oxylipin profiling reveals the preferential stimula- G., De Wit, P. J. G. M. & Joosten, M. H. A. J. No evi- 19846001 C2 (2000). Elicitor Pep-13 aus Phytophthora sojae. University expression in response to herbivore damage in tion of the 9-lipoxygenase pathway in elicitor-trea- dence for binding between resistance gene product Clemens, S., Thomine, S. & Schroeder, J. I. Molecular of Halle-Wittenberg, Department of Biology, Arabidopsis thaliana. Physiol. Plant. 114, 85-91 ted potato cells. J. Biol. Chem. 276, 6267-6273 Cf-9 of tomato and avirulence gene product AVR9 mechanisms that control plant tolerance to heavy 17/08/2001. (2002). (2001). of Cladosporium fulvum. Mol. Plant Microbe metals and possible roles towards manipulating Doctoral Theses Interact. 14, 867-876 (2001). metal accumulation. In: Plant Biotechnology and Bau, Stephan: Untersuchungen zur Jasmonat- Haase, Stefanie: Untersuchung putativer Schwer - Bloss, T., Clemens, S. & Nies, D. H. Characterization Göbel, C., Feussner, I., Hamberg, M. & Rosahl, S. Transgenic Plants (Oksman-Caldentey, K.-M. & Signaltransduktion in Arabidopsis thaliana anhand metalltransporter aus Schizosaccharomyces of the ZAT1p zinc transporter from Arabidopsis Oxylipin profiling in pathogen-infected potato lea- Nass, N. & Scheel, D. Enhanced luciferin entry cau- Barz, H.W., eds.) Marcel Dekker, Inc., New York, pp. des Jasmonat-regulierten Gens Atjrg21. University pombe. University of Halle-Wittenberg, thaliana in microbial model organisms and reconsti- ves. Biochim. Biophys. Acta 1584, 55-64 (2002). ses rapid wound-induced light emission in plants 665-691 (2002). of Halle-Wittenberg, Department of Department of Biochemistry/Biotechnology, tuted proteoliposomes. Planta 214, 783-791 (2002). expressing high levels of luciferase. Planta 212, 149- Biochemistry/Biotechnology, 26/01/2001. 28/01/2002. Hornung, E., Rosahl, S., Kühn, H. & Feussner, I. 154 (2001). Hirt, H. & Scheel, D. Receptor-mediated MAP kina- Blume, B., Nürnberger, T., Nass, N. & Scheel, D. Creating lipoxygenases with new positional specifi- se activation in plant defense. In: Results and De Figueiredo, Clarice: Physiologisch-biochemische Landtag, Jörn: Transformation von Kartoffel- und Receptor-mediated increase in cytoplasmic free cities by site-directed mutagenesis. Biochem. Soc. Neumann, D. & De Figueiredo, C. A novel mecha- Problems in Cell Differentiation, Vol. 27. MAP Mechanismen der Schwermetalltoleranz bei Tabakpflanzen mit der Tyrosin-Decarboxylase-2 calcium required for activation of pathogen defense Trans. 28, 825-826 (2000). nism of silicon uptake. Protoplasma 220, 59-67 Kinases in Plant Signal Transduction (Hirt, H., ed.) Armeria mari tima (Mill.) Willd. ssp. halleri (Wallr.). cDNA aus Petersilie und Untersuchung der in parsley. Plant Cell 12, 1425-1440 (2000). (2002). Springer-Verlag, Heidelberg, pp. 85-93 (2000). University of Halle-Wittenberg, Department of Expression des Transgens. University of Halle- Ichimura, K., Shinozaki, K., Tena, G., Sheen, J., Henry, Y., Biochemistry/Bio technology, 24/09/2002. Wittenberg, Department of Biology, April 2001. Brunner, F., Rosahl, S., Lee, J., Rudd, J. J., Geiler, C., Champion, A., Kreis, M., Zhang, S., Hirt, H., Wilson, C., Neumann, D. & zur Nieden, U. Silicon and heavy Scheel, D. Parasitismus im Pflanzenreich. In: Kauppinen, S., Rasmussen, G., Scheel, D. & Heberle-Bors, E., Ellis, B. E., Morris, P. C., Innes, R. W., metal tolerance of higher plants. Phytochemistry Parasitismus als Lebensform. Nova Acta Leopoldina 316 Fellbrich, Guido: Interaktionen zwischen Pflanzen Simm, Claudia: Phytochelatinsynthase aus Schizo - Nürnberger, T. Pep-13, a plant defense-inducing Ecker, J. R., Scheel, D., Klessig, D. F., Machida,Y., Mundy, J., 56, 685-692 (2001). NF. , Nr. 83, Barth, Heidelberg, S. 25-31 (2000). und phytopathogenen Oomyceten. Isolierung, saccharomyces pombe. Untersuchungen zur pathogen-associated pattern from Phytophthora. Ohashi, Y. & Walker, J. C. Mitogen-activated protein Sequenzierung und partielle Charakterisierung Lokalisierung, Regulation und biochemischer EMBO J. 21, 6681-6688 (2002). kinase cascades in plants: a new nomenclature. Trends Newman, M.-A., von Röpenack-Lahaye, E., Parr, A., Scheel, D., Blume, B., Brunner, F., Fellbrich, G., eines Proteinelicitors aus Phytophthora parasitica. Funktionalität. University of Halle-Wittenberg, Plant Sci. 7, 301-308 (2002). Daniels, M. J. & Dow, J. M. Induction of hydroxycin- Dalboge, H., Hirt, H., Kauppinen, S., Kroj, T., University of Halle-Wittenberg, Department of Department of Biochemistry/Biotechnology, Brunner, F., Wirtz, W., Rose, J. K. C., Darvill, A. G., namoyl-tyramine conjugates in pepper by Ligterink, W., Nürnberger, T., Tschöpe, M., Zinecker, Biochemistry/Biotechnology, 08/07/2001. 12/07/2000. Govers, F., Scheel, D. & Nürnberger, T. A ß-glucosida- Kamphausen, T., Fanghähnel, J., Neumann, D., Schulz, Xanthomonas campestris, a plant defense response H. & zur Nieden, U. Receptor-mediated signal se/xylosidase from the phytopathogenic oomycete, B. & Rahfeld, J.-U. Characterization of Arabidopsis activated by hrp gene-dependent and hrp gene- transduction in plant defense. In: Biology of Plant- Göbel, Cornelia: Untersuchungen zur Funktion von Tennstedt, Pierre: Untersuchungen zur Abwehr - Phytophthora infestans. Phytochemistry 59, 689- thaliana AtFKBP42 that is membrane bound and independent mechanisms. Mol. Plant Microbe Microbe Interactions, Vol. 2 (de Wit, P. J. G. M., Oxylipinen bei der Pathogenantwort in Solanum expression in Arabidopsis-Mutanten. University of 696 (2002). interacts with HSP90. Plant J. 32, 263-276 (2002). Interact. 14, 785-792 (2001). Bisseling, T. & Stiekema, W. J., eds.) International Society for Molecular Plant-Microbe Interactions, tuberosum L. University of Halle-Wittenberg, Halle-Wittenberg, Department of Biochemistry/ Department of Biochemistry/Biotechnology, Biotechnology, 29/07/2002. < Bruns, I., Sutter, K., Menge, S., Neumann, D. & Krauss, Kroj, T., Rudd, J. J., Nürnberger, T., Gäbler, Y., Lee, J. & Newman, M.A., von Roepenack-Lahaye, E., Parr, A., St. Paul, pp. 131-135 (2000). 05/12/2001. G.-J. Cadmium lets increase the glutathione pool in Scheel, D. Mitogen-activated kinases play an essen- Daniels, M. J. & Dow, J. M. Prior exposure to lipopo- bryophytes. J. Plant Physiol. 158, 79-89 (2001). tial role in oxidative burst-independent expression lysaccharide potentiates expression of plant defen- Scheel, D. Oxidative burst and the role of reactive Kemmerling, Birgit: Identifizierung und Charak - of pathogenesis-related genes in parsley. J. Biol. ses in response to bacteria. Plant J. 29, 487-495 oxygen species in plant-pathogen interactions. In: terisierung systemisch responsiver Gene der Cazalé, A.-C. & Clemens, S. Arabidopsis thaliana Chem. published November 7, 2002 as (2002). Oxidative Stress in Plants (Inzé, D. & van Montagu, Kartoffel (Solanum tuberosum L.) nach Inokulation expresses a second functional phytochelatin syn- 10.1074/jbc.M208200200. M., eds.) Taylor & Francis, London, pp. 137-153 mit dem nichtpathogenen Bakterium Pseudomonas thase. FEBS Lett. 507, 215-219 (2001). Noeringer, C., Scheel, D. & Blee, E. Lipoxygenase (2002). syringae pv. maculicola. University Halle- Landgraf, P., Feussner, I., Hunger, A., Scheel, D. & Rosahl, isoforms in elicitor-treated parsley cell suspension Wittenberg, Department of Biology, 06/09/2001. Clemens, S. Molecular mechanisms of plant metal S. Systemic accumulation of 12-oxo-phytodienoic acid cultures. Biochem. Soc. Trans. 28, 2827-2829 Scheel, D. Signal transduction elements. In: Plant tolerance and homeostasis. Planta 212, 475-486 in SAR-induced potato plants. Eur. J. Plant Pathol. 108, (2000). Biotechnology and Transgenic Plants (Oksman- Lubaretz, Olga: Non-stress induced small head (2001). 279-283 (2002). Caldentey, K.-M., Barz, H. W., eds.) Marcel Dekker, shock proteins in higher plants. University of Halle- Nürnberger, T. & Scheel, D. Signal transmission in Inc., New York, pp. 427-444 (2002). Wittenberg, Department of Clemens, S. Developing tools for phytoremediation: Landtag, J., Baumert, A., Degenkolb, T., Schmidt, J., the plant immune response. Trends Plant Sci. 6, Biochemistry/Biotechnology, 14/06/2001. Towards a molecular understanding of plant metal Wray, V., Scheel, D., Strack, D. & Rosahl, S. 372-379 (2001). Scheel, D. & Wasternack, C., eds. Plant Signal tolerance and accumulation. Int. J. Occup. Med. Accumulation of tyrosol glucoside in transgenic Transduction., Oxford University Press, Oxford, Patzlaff, Astrid: Untersuchungen zur Expression von Environm. Health 14, 235-239 (2001). potato plants expressing a parsley tyrosine decar- Nürnberger, T. & Brunner, F. Innate immunity in (2002). Peroxidase Ca aus Arabidopsis thaliana und boxylase. Phytochemistry 60, 683-689 (2002). plants and animals: emerging parallels between the Generierung von Mutanten mit veränderter Clemens, S., Bloss, T., Vess, C., Neumann, D., Nies, D. recognition of general elicitors and pathogen-asso- Scheel, D. & Wasternack, C. Signal transduction in Expression. University of Halle-Wittenberg, H. & zur Nieden, U. A transporter in the endoplas- Lee, J., Klessig, D. F. & Nürnberger, T. A harpin bin- ciated molecular patterns. Curr. Opin. Plant Biol. 5, plants: cross talk with the environment. In: Plant Department of Biochemistry/Bio technology, mic reticulum of Schizosaccharomyces pombe cells ding site in tobacco plasma membranes mediates 318-324 (2002). Signal Transduction (Scheel, D., Wasternack, C., 22/06/2001. mediates zinc storage and differentially affects tran- activation of the pathogenesis-related gene HIN1 eds.) Oxford University Press, Oxford, pp. 1-5 sition metal tolerance. J. Biol. Chem. 277, 18215- independent of extracellular calcium but depen- Petters, J., Göbel, C., Scheel, D. & Rosahl, S. A patho- (2002). Petters, Julia: Isolierung und Charakterisierung 18221 (2002). dent on mitogen-activated protein kinase activity. gen-responsive cDNA from potato encodes a pro- Books and Book chapters in press pathogen- und stressresponsiver Gene der Plant Cell 13, 1079-1093 (2001). tein with homology to a phosphate-starvation Kartoffel (Solanum tuberosum L.). University of Clemens, S., Palmgren, M. G. & Krämer, U. A long induced phosphatase. Plant Cell Physiol. 43, 1049- Clemens, S., Simm, C. & Maier, T. Heavy metal bin- Halle-Wittenberg, Department of Pharmacy, way ahead: understanding and engineering plant Lee, J., Klüsener, B., Tsiamis, G., Stevens, C., Neyt, C., 1053 (2002). ding proteins and peptides. In: Biopolymers, Vol. 7 23/11/2001. metal accumulation. Trends Plant Sci. 7, 309-315 Tampakaki, A. P., Panopoulos, N. J., Nöller, J., Weiler, E. Polyamides and complex proteinaceous Materials (2002). W., Cornelis, G. R., Mansfield, J. W. & Nürnberger, T. Stumpe, M., Kandzia, R., Göbel, C., Rosahl, S. & Part A, (Fahnestock, S. R., ed.) Wiley-VCH, New York (2003). Romanski, Annette: Das Elicitorprotein NPP1 - HrpZPsph from the plant pathogen Pseudomonas Feussner, I. A pathogen-inducible divinyl ether syn- Isolierung und Charakterisierung der korrespon- Clemens, S., Schroeder, J. I. & Degenkolb, T. syringae pv. phaseolicola is exported by the type III thase (CYP74D) from elicitor-treated potato dierenden cDNA, heterologe Expression des Caenorhabditis elegans expresses a functional phy- secretion pathway and forms an ion-conducting pore suspension cells. FEBS Lett. 507, 371-376 (2001). Lee, J. & Nürnberger, T. Pseudomonas syringae Proteins und Studien zur Signalperception. tochelatin synthase. Eur. J. Biochem. 268, 3640-3643 in vitro. Proc. Natl. Acad. Sci. U.S.A., 98, 289-294 pathovars and related pathogens. In: Developments University of Halle-Wittenberg, Department of (2001). (2001). in Plant Pathology, Vol. 10, (Mansfield, J.W. & Vivian, Dow, M., Newman, M.-A. & von Roepenack, E. The Lee, J. & Rudd, J. J. Calcium-dependent protein kina-
54 55 Department: Secondary Metabolism Head: Prof. Dieter Strack Secretary: Heidemarie Stolz
ork of the department is con- droxycinnamoyltransferases Wcerned with the molecular as well as several hydroxy- regulation of plant secondary meta- cinnamate glucosyltrans- bolism, evolution of the enzymes ferases from A ra bidopsis thali- involved in the biosynthesis of secon- ana and ra pe (Bras si ca na pus). dary products and their role in inter- In addition flavonoid and actions of plants with their environ- betanidin glucosyltransfera- ment. ses from be tacyanin- accumulating plants or fla- The work on metabolic regulation vonoid methyltransferases includes isolation and characteriza- from the ice plant (Me sem - tion of the corresponding enzymes bryan themum crys tallinum) and the encoding genes, focu- are investigated. sing on transferases. We cur - rently investigate ma late The aim of the work on glu- and choline hy - cosyl- and hy - droxycinnamoyltransferases is to elu- jasmonates, in development and func- cidate their evo lutionary origin and tional mainte-nance of mycorrhizal structure-function re lations to pre- symbiosis. These studies are suppor- dict substrate specificity. Glu cosyl - ted by comprehensive analysis of pri- trans ferases in vol ved in betacyanin mary and secondary metabo-lites biosynthesis are considered to be ("metabolite profiling") in wild-type oligophyletic and originate from dif- and transgenic mycorrhizal plants. < ferent clusters of flavonoid glucosyl- transferases. Hydroxy cinna moyl- trans fe ra ses, which are dependent on b-acetal esters as acyldonors, are vacuolar serine carboxypeptidase- like (SCPL) proteins as found for the enzyme involved in the formation of sinapoylmalate in Ara bidopsis. The general existence of vacuolar b-ace- tal ester-de pen dent acyltransferases would prove a new concept of cell com partmentation of plant se- condary metabolism.
Special emphasis is also placed on programs focusing on the molecular interactions of plants with arbuscular mycorrhizal fungi. The work of two groups is concerned with fungus- induced alterations in plant isopre- noid metabolism, in particular caro- tenoid biosynthesis and degradation, accompanied by a dramatic reorgani- zation of plastid population in arbu- scule-harbouring root cells. Another main objective is the analysis of the role of phytohormones, in particular
56 57 Research Group: Molecular Physiology of Mycorrhiza Head: Michael H. Walter
Group Members Most herbaceous plants form symbiotic associations with a small number of with Glo mus mosseae and G. intraradi- tein from E. coli has been used to create Thomas Fester fungi in the rhizosphere in order to improve their water uptake and acquisi- ces. For the first time in plants the exi- specific antibodies. Their use in immuno- (postdoctoral position until December 2001) tion of mineral nutrients. These interactions are called arbuscular mycorrhizas stence of two distinct, only distantly rela- localisation studies vi - Kristine Halfmann (AM), a term derived from the haustoria-like fungal arbuscules developing in ted classes of DXS genes could be dedu- sua lized an ex - (PhD student until February 2000) the root cortex. The work of the group focuses on alterations in plant isopre- ced from the analysis of M. truncatula ten sive net- Joachim Hans cDNAs. Only the expression of DXS2 work of in - (PhD student) noid metabolism induced by AM fungi, in particular on reactions located in pla- from M. truncatula is regulated by arbu- Swanhild Lohse stids. Starting from metabolite analyses of various apocarotenoids, a number scular mycorrhizal fun gi. DXS1 is expres- (PhD student until December 2001) of fungus-stimulated gene activities from their biosynthetic pathway could be Kerstin Manke sed in most tissues at a constitutive level (technician) characterized. These include steps from the non-mevalonate methylerythritol and appears to fulfill mainly housekee- Alexander Röhrig phosphate (MEP) and from the carotenoid pathways. For the first reaction of ping functions. Similar complementary terconnected (student since August 2002) the MEP pathway a diversification and specific expression of a 1-deoxy-D-xylu- expression profiles and mycorrhiza- plas tids a round ma - Sudha Sahay lose 5-phos-phate synthase 2 (DXS2) gene in mycorrhizal roots was shown.A regulation of DXS2 transcript levels ture fungal arbuscules in (DAAD-fellow until August 2001) new concept of dedicated roles of DXS1 and DXS2 in the biosynthesis of pri- were found in maize, tomato and tobac- colonized cortical cells. The most recent Michael Stephan (postdoctoral position until September 2001) mary and secondary isoprenoids has been introduced. co. Additional data suggest an involve- work involves the carotenoid cleavage Gerlinde Waiblinger ment of DXS2 in the biosynthesis of step performed by carotenoid cleaving (technician until December 2001) many other secondary isoprenoids such dioxygenases (CCDs). Several cDNA as leaf trichome monoterpenes of mint clones from both M. truncatula and Metabolite analysis of roots of various various plant families could be shown. and solanaceous species, petal caroteno- maize have been isolated. Initial analyses Collaborators plants including cereals, tobacco and The apocarotenoids are presumably in - ids, and terpenoid indole alkaloids. As a in maize indicate a mycorrhiza-media- Jörg Degenhardt legumes colonized by the mycorrhizal tegrated into a complex mixture of es - result, a new concept of dedicated DXS ted regulation of this step as well. Max Planck Institute for Chemical Ecology, Jena, Germany fungus Glomus intraradices has led to ters between mycorradicin and glycosy- enzymes for primary and secon-dary iso- Philipp Franken Max Planck Institute for Terrestrial Microbiology, the identification of two classes of apo- lated C13 cyclohexenone derivatives. Ac - prenoids can be introduced (see figure). Another project targeted more Marburg, Germany carotenoids: (i) glycosylated C13 cyclohe- cumulation of this complex in mycorrhi- Genomic sequences harbouring DXS2 general aspects of plastid develop- Giovanni Giuliano xenone derivatives and (ii) an acyclic C14 zal roots correlates with degradation of genes have been isolated. These materi- ment during the symbiosis. Use Ente per le nuove tecnologie, l energia e l ambiente, ENEA, Rome, Italy po ly ene compound termed mycorradi- fungal arbuscules. als will provide useful tools for gene sup- of transgenic tobacco plants ex pres sing a Bettina Hause, Jürgen Schmidt cin. Further biochemical work has now pression studies and promoter analyses. plastid-directed green fluorescent pro- Institute of Plant Biochemistry, Halle, Germany provided a facile and sensitive detection Early steps of apocarotenoid biosynthe- tein has shown dramatic changes in Phylogenetic tree of DXS proteins from plants and the photosynthetic bacterium Rhodobacter capsula- Martin Parniske method for mycorradicin. The wide- sis are catalyzed by the enzymes 1-deoxy- A single class of cDNAs for DXR has mycorrhizal roots with a similar network tus. Branches underlined in green indicate preferenti- John Innes Center, Norwich, UK spread but not universal occurrence of D-xylulose 5-phosphate synthase (DXS) been isolated from a maize mycorrhizal of plastids around arbuscules as seen al expression in green tissues. Conversely, orange back ground indicates expression correlated with the Andreas Perlick this compound in mycorrhizal roots of and 1-deoxy-D-xylulose 5-phosphate re - root library. DXR transcript levels are with the DXR antibody. These networks biosynthesis of secondary isoprenoids such as apoca- University of Bielefeld, Germany duc toisomerase (DXR) elevated in mycorrhizal maize roots but are highly dynamic structures appearing rotenoids of mycorrhizal roots, petal carotenoids or Ajit Varma monoterpenes of leaf trichomes. The data introduce Jawaharlal Nehru University, New Delhi, India as part of the re - not to the high extent as has been found and disappearing concomi-tantly with a concept of dedicated roles of DXS1 and DXS2 in primary and secondary functions, respectively. Victor Wray cently discovered for DXS2. Recombinant maize DXR pro- formation and degradation of arbuscules German Research Centre for Biotechnology, MEP pathway loca- (see report B. Hause). < Braunschweig, Germany ted to plastids. Eleonore Wurtzel City University New York, Bronx, USA Strongly elevated trans cript levels for both enzymes com - pared to controls were shown for my - corrhizal roots of se veral cereals. A more detailed ana- lysis of DXS gene regulation and or - ga nization was ini - ti ated for the mo - Visualization of plastid reorgani - zation in a mycorrhized root cell. Isoprenoid biosynthesis and its compartmentation. Two separate del legume Medi - A fluorescence-labeled antibody pathways lead to the key intermediate isopentenyl diphosphate cago truncatula specific for DXR reacts with pla- (IPP). The apocarotenoids blumenin (C13 cyclohexenone derivative) stids covering a fungal arbuscule and mycorradicin accumulating in mycorhizal roots are highlighted. and its interaction (right) or surrounding a nucleus (upper left).
58 59 Research Group: Cell Biology of Mycorrhiza Head: Bettina Hause
Group Members Plant hormones are believed to play a role in the establishment and develop- osmotic stress or in defense against biotic biosynthesis in AM roots, we have shown Thomas Fester ment of symbiotic interactions between plants and arbuscular fungi (arbuscu- stresses takes place. As a consequence, that this process is virtually ubiquitous in (leader junior group since January 2002) lar mycorrhiza, AM). Jasmonates, known as regulators in stress responses of mycorrhizal roots may be more resistant the plant kingdom. The extent of activa- Ulrike Hintsche plants against various biotic and abiotic stresses, might be important regula- against secondary infection and/or osmotic tion, however, is variable regarding the (technician) tors of this symbiosis. Therefore, the main objective of our group is the ana- stresses. respective plant species. One major cha- Stanislav Isayenkov racteristic of the phenomenon is the fin- (PhD student until October 2002, lysis of the role of jasmonates during the interaction between Glomus intra- afterwards postdoctoral position) radices and barley (Hordeum vulgare) or barrel medic (Medicago truncatula). To test this hypothesis we intend to per- ding that carotenoid intermediates of the form functional analyses by modulating jas- pathway are present only in very small Swanhild Lohse In a second project, the activation of carotenoid biosynthesis in AM roots is (PhD student since January 2002) monate content in mycorrhizal roots of M. amounts, even in plants accumulating large Tamás Monostori studied. This activation is connected with a massive proliferation of the pla- truncatula. A cDNA coding for the allene amounts of apocarotenoids (mycorradicin (PhD student until March 2001) stids of colonized root cortical cells. Cell biological phenomena as well as oxide cyclase (AOC), the enzyme perfor- and glycosylated cyclohexenone derivati- Constantin Rüder underlying molecular changes of the plastid proliferation will be elucidated. (student until December 2000) ming the crucial step in JA biosynthesis, ves). Two hypothetical functional reasons Sara Schaarschmidt was isolated from M. truncatula. Vectors for the activation of carotenoid biosynthe- (PhD student since May 2002) were constructed containing this cDNA in sis in AM roots are currently investigated: Diana Schmidt sense or antisense orientation or the (i) The possible induction of carotenoid (student until July 2001) A possible role of jasmonates in the in situ-techniques (in situ-hybridization, RNAi construct, all of them under control biosynthesis by reactive oxygen species Carola Tretner mycorrhizal interaction is indicated by the im munocytochemistry) accumulation of of the CaMV35S promoter. After transfor- (ROS) produced during establishment of (research scientist since September 2002) following data: Jasmonic acid (JA), applied AOS and JIP23 mRNA and protein, respec- mation, plants are expected with increa- the AM symbiosis and a possible protec- Gerlinde Waiblinger (technician since January 2002) exogenously, promotes colonization and tively, could be shown to occur in cells sed endogenous levels of JA (AOCsense) tion of the plant cell against such ROS by de velopment of mycorrhizal or de creased levels of JA (AOCantisense, carotenoids; and (ii) a possible metabolic struc tures, and the endogenous AOC-RNAi). Assuming that altered JA- role of the chlororespiratory activity Collaborators JA level of mycorrhizal roots is levels lead to altered mycorrhizal phenoty- involved in carotenoid biosynthesis. remarkably higher than that of pes, cell biological and biochemical appro- Peter Bramley, Paul Fraser non-mycorrhizal roots. The in - aches will be used to analyze these pheno- The accumulation of apocarotenoids in University of London, UK crease of JA content in barley types. Additionally, gene expression studies AM roots might be part of a complex Ivo Feussner, Uwe Sonnewald Institute of Plant Genetics and Crop Plant Research, roots upon mycorrhization is will be performed using cDNA microar- reorganisation of plastid structure and Gatersleben, Germany accompanied by the expression rays pro vided by the DFG Research Focus me tabolism in AM roots. Root cortical Philipp Franken of genes coding for enzymes of Pro gram 1084 "Molecular Basis of Mycor - cell plastids are responsible for a number Max Planck Institute for Terrestrial Microbiology, Marburg, Germany JA biosynthesis (allene oxide rhizal Symbioses". In a second approach, of biosynthetic processes, which are Giovanni Giuliano synthase, AOS) and for jasmona- transgenic tobacco plants were used, essential for the formation and functio- Ente per le nuove tecnologie, l energia e l ambiente, ENEA, Rome, Italy te-induced proteins (JIP23). In which express a yeast invertase targeted ning of the symbiotic interface. In accor- Gerd Hause order to record the kinetics of to the apoplast. These plants exhibit alte- dance with this functional importance, University of Halle, Germany JA accumulation during develop- red source-sink relationships and will be massive proliferation of plastids in colo- Willy Peumans, Els Van Damme ment of mycorrhizal structures, analyzed with respect to alterations in the nized tobacco root cortical cells leading University of Leuven, Belgium a system of "near-synchronous" mycorrhizal phenotype expected on bio- to network-like structures covering the Thomas Roitsch mycorrhization was established chemical, molecular and cytological level. arbuscules have been observed. Currently, University of Würzburg, Germany by the use of nurse-pot cultures. From both approaches, we hope to get we are analyzing changes in the expres- Claus Wasternack, Jürgen Schmidt, Confocal laser scanning micrographs of tobacco Since JA levels increase later than the ini- harboring arbuscules (see figure). From all insights into the relationship of mycor- sion levels of plastid-related genes using root cortex non-colonized (A) and colonized (B, C) Otto Miersch by an AM fungus, respectively. The plastids are visu- Institute of Plant Biochemistry, Halle, Germany tial steps of the plant-fungal interaction data ob-tained, the following hypothetical rhiza, jasmonate action and the sugar sta- DNA-arrays and Real-Time RT-PCR. alized by the green fluorescent protein targeted to Victor Wray occur, the development of mycorrhiza rather scenario is suggested: The plant root sup- tus within the mycorrhizal roots. These analyses will provide first infor- plastids (transgenic tobacco plants courtesy of M. Hanson, Ithaca, New York, USA). In colonized cells German Research Centre for Biotechnology, than the recognition of the interacting plies the fungus with carbohydrates Þ the mation regarding the molecular and bio- Braunschweig, Germany the plastids formed a network-like structure partners may cause expression of JA-bio- plant root becomes a stronger sink organ Concerning the activation of carotenoid chemical changes underlying the process around the arbuscules. Bars represent 50 µm in A, synthetic genes and finally elevate JA upon mycorrhization resulting in an B, and 25 µm in C. levels. In addition to the temporal pattern, enhanced accumulation of soluble sugars the spatial pattern of gene expression ap - within the apoplast Þ expression of genes pearing during the development of the coding for enzymes of JA biosynthesis fungal organs within the root cortex (vesi- occurs Þ level of jasmonates increases Þ cles, arbuscules) was recorded. By use of induction of genes involved in response to
Figure: In situ-localization of AOS-transcripts within mycorrhizal barley roots. The detection performed Somatic regeneration of Medicago truncatula. Plant explants give with the antisense probe (A, B) exhibits a clear staining of the cytoplasm of root cortex cells containing rise to embryogenic callus (A), which then develops small embryos fungal structures (arrow head in B), whereas the negative control (sense, C, D) does not show labeling within (B). After transfer of embryos to "Embryo-Developing-Media" small arbuscule-containing cells (arrow in C, arrow head in D). Bars represent 50 µm. plantlets are formed (C).
60 61 Research Group: Biochemistry of Mycorrhiza (since 2002) Head: Willibald Schliemann
Group Members The research is focused on the comprehensive analysis of alte-rations of pri- the levels are different for the individual Christian Ammer mary and secondary metabolite patterns during the establishment of the com ponents. In cooperation with the (research scientist since September 2002) arbuscular mycorrhizal symbiosis in the model system Medicago truncatula / working group of Thomas Fester (IPB) Barbara Kolbe Glomus intraradices with the aim to characterize the causal relationships bet- mycorrhiza-induced cyclohexenone de ri - (technician) ween gene expression and metabolite profiles during the symbiosis. Metabolic vatives were detected in roots of Zea Lars Seipold mays, Medicago truncatula and Orni tho - (PhD student since June 2002) processes that are essential for the functioning of this root-fungus system galum umbellatum by HPLC. In the latter have to be elucidated that may be of general importance also in other mycor- material ( see figure), besides the "yellow rhizal systems. Furthermore, metabolite analysis of transgenic M. truncatula Collaborators pigment" and cyclohexenones, a group of plants is intended to evaluate the effect of gene transfer or knockouts on the hitherto unknown apocarotenoids with Thomas Degenkolb, Bettina Hause, kinetics of mycorrhizal symbiosis and phenotypical changes in plant develop- spectral properties similar to mycorradi- Thomas Fester, Jürgen Schmidt, Michael H. Walter ment (in cooperation with projects of the DFG Research Focus Program cin was observed suggesting that they Institute of Plant Biochemistry (IPB), Halle, Germany 1084 "Molecular Basics of Mycorrhizal Symbioses"). might be precursors of the "yellow pig- Philipp Franken ment". Using lecaton from a leek/G. Max Planck Institute for Terrestrial Microbiology, Marburg, Germany intraradices preculture for M. truncatula Inna Kuzovkina inoculation a fast and efficient mycorrhi- Timiryasev Institute of Plant Physiology, Russian RP-HPLC-PDA, LC-ESI-MS and GC- stans) and GC-MS (amino acids, aliphatic zation (ca. 90 % after 4 weeks) was Academy of Sciences, Moscow, Russia TOF-MS are the methods used in our acids, phenylpropanoids, in particular iso- observed which will be used in detailed Karsten Niehaus University of Bielefeld, Germany metabolite profiling approach. At the flavonoids, sugars, sterols) to facilitate studies of mycorrhization kinetics. Manfred Nimtz, Victor Wray beginning databases of reference com- subsequently the dereplication of the German Research Centre for Biotechnology, pound were created using HPLC (flavo- endogenous compounds. To reduce the To achieve an adequate handling of the Braunschweig, Germany The legume barrel medic (Medicago truncatula Gaertn. cv. Jemalong A17), the model plant noids, isoflavonoids, pterocarpans, cou- chemical complexity of the metabolome, quantitative metabolite profiling data, an for functional genomic approaches to arbuscular mycorrhizal symbiosis. mestans and their glucosides), LC-MS sequential extractions of lyophilized root efficient computing system with statistic (isoflavonoids, pterocarpans, coume- material with dichloromethane, acetone software was recently installed. In coor- and 80% aqueous methanol were dination with other groups of the institu- performed. In these extracts my - te dealing with similar bioinformatic pro- cor rhiza-specific alterations of more blems, these tools will be used for valida- than 300 root metabolites were tion, statistical evaluation and meaningful ob served by HPLC. In LC-MS the presentation of the results. In the future accumulation of different isoflavo- material from transgenic plants provided noid glucosides and their corre- by the collaborating groups will be analy- sponding malonates as well as sapo- zed to determine the effect of the gene- nins was detected. By GC-MS of tic alterations on the metabolite pattern the dichloromethane extract of and to correlate the metabolite profiles twelve weeks old mycorrhizal roots with the gene ex pression profiles in a (M. truncatula / G. intraradices - 40 functional genomic approach. < % mycorrhization) a dramatic incre- ase of palmitelaidic and oleic acid was detected, whereas other long- chained fatty acids decreased in comparison to non-mycorrhizal controls. In all ex tracts the levels of some acids of the primary metabo- lism (lactic, malic, malonic, succinic, citric, g-amino butyric and trihydro- xybu-tyric acids) were higher in Arbuscules of Glomus intraradices in the barrel medic root (trypan blue staining). non-mycorrhizal than in mycorrhi- zal roots. Sugars and inositol deri- Star-of-Bethlehem (Ornithogalum umbellatum L., Hyacinthaceae) va-tives are the predominating Top: flowering plants; bottom: roots after mycorrhization (6 compounds in the acetone and 80 months) with Glomus intraradices in comparison to non- mycorrhized controls (micrographs by courtesy of T. Fester, % aqueous methanol extracts, but
62 63 Research Group: Glycosyltransferases Head: Thomas Vogt
Group members Based on earlier work, our group started and 6-GT from Dorotheanthus bellidi- Stabilisation and solubilisation of plant natural products are performed by a specificity. No transcripts en coding Stefan Ebert (student until July 2000) wide range of glucosyltransferases (GTs) with often overlapping substrate GTs were found. Among Mwafaq Ibdah specificities. These enzymes may also detoxify bioactive low-molecular weight Besides our approach to correlate GT a va riety of induced (PhD student until August 2002) compounds, in particular those from exogenous sources. Sequence identities function with sequence information, our transcripts ranging from Judith Hans research also contributes to the investi- catalase to JIP-23 (PhD student since Mai 2000) cluster GTs according to regiospecificities rather than substrate specificities imply that this superfamily of proteins has evolved oligophyletically as one of gation of betacyanin biosynthesis in the (Jasmonat Induced Pro - Dagmar Knöfel Caryophyllales. Several lines of evidence, tein) or a salt to lerant (technician) the primary adaptive mechanisms of plants to meet the changing environmen- including cloning of highly homologous pro tein, several O-me - Ute Vinzens tal conditions in a timely and developmentally controlled manner. Similar obser- (technician until September 2002) 5- and 6-GT sequences from red beet thyl transferases (OMTs) vations hold true for the superfamily of plant O-methyltransferases (OMTs), (Beta vulgaris), suggest a conserved gly- were selected as putati- with a new subclass of enzymes involved in the modification of UV-induced cosylation of betacyanins at the betani- ve candidates to be in - Collaborators flavonoid conjugates discovered recently. din level among all families within the volved in the methy- Hans Bohnert Caryophyllales. GTs performing either lation of the ob served University of Urbana, Illinois, USA 5- or 6-glycosylation are phylogeneti- (UV-) light-induced flavo- John Cushman cally derived from two different classes nol conjugates. One of the correspon- Red coloration of the ice plant due to epidermal University of Reno, Nevada, USA on the molecular physiology of betalains, formis (Dbs), we were able to demon- of enzymes, involved in the position- ding OMT-proteins showed the requi- accumulation of betacyanins and flavonol conju - gates after five days of exposure to high light (1500 Patrik Jones 2 Chiba University, Chiba, Japan but lately as a result of the ongoing strate an oligophyletic origin of the cor- specific glycosylation of flavonoids or red enzyme activities, and its presence µM/m x s) irradiation is observed only for plant A work, the major focus has shifted responding glucosyltransferase genes other hydroxylated phenylpropanoids. was consistent with the occurrence of (UV-A/B radiation, cut-off filter 305 nm), but not for Toni M. Kutchan, Sabine Rosahl, plant B (UV-A/B radiation, cut-off filter 360 nm). Jürgen Schmidt towards enzymes involved in the modifi- from different clusters of flavonoid GTs. The presence of a GT, glycosylating the conjugated flavonol-6,3 -di-O- Institute of Plant Biochemistry, Halle, Germany cation of the two classes of plant natural From this study, it is concluded that cyclo-dopa, which has been proposed as methylether derivatives in light-induced Vladimir Kuznetsov, Inna Kuzovkina products under investigation, the betacy- regiospecificity and not substrate specifi- the glucose acceptor, cannot be ruled bladder cells of the ice plant. This pro- Timiryazev Institute of Plant Physiology, Russian Academy of Sciences, Moscow, Russia anins and the flavonoids. Primarily, our city ap pears to be the organizing princi- out, but is most unlikely. tein was purified from leaves of the ice Ullrich Matern work has been directed towards a de- ple in cluster formation. This may have plant. Based on amino acid sequence University of Marburg, Germany tailed understanding of the biochemistry important consequences for evaluating Our second model system, the ice information, the cDNA was cloned Harald Seidlitz, Werner Heller and molecular evolution of plant natural the structure/function relationship wit- plant (Mesembryanthemum crystalli- from a cDNA library and ex pressed in National Research Center for Environment and Health, Munich, Germany product glycosyltransferases. By detailed hin the rapidly growing genomic databa- num), is faced with extreme arid condi- a prokaryotic system. The recombinant sequence analysis and substrate specifi- ses for a variety of crop and non-crop tions in its na-tural habitat. Besides its enzyme displayed high position-specifi- city studies of two GTs, the betanidin 5-GT species, like rice, corn or Arabidopsis, well-studied adap tation to drought city towards me thy lation of ortho-dihy- where a total of 110 GT-sequences with stress, the plant is capable of tolerating droxyl groups, with the ability to largely unknown substrate specificities exposure to ex-treme light intensities, methylate a variety of potential sub- has al-ready been described. Site-direc- combined with a high dose of UV radi- strates, including flavo-noids, hydroxy- ted mu ta genesis, performed with the ation, by a rapid ac cumulation of glyco- cinnamic acids and their corresponding heterologously expressed 5-GT protein sylated and methylated flavonol and CoA-esters. Protein sequence analysis from Dbs, among more than 20 conser- betacyanin conjugates in leaf epidermal indicates that this flavo noid-methylating ved residues, indicate the presence of layers (see figure). At the molecular activity most likely defines a new sub- several catalytically essential amino acid level a subtractive cDNA library of a group of small, Mg2+-dependent OMTs, resi dues. They are probably involved in light-in-duced versus non-induced lea- previously shown to be involved only in the substrate binding of all GTs of the b- ves revealed the presence of several the methylation of the lignin precursor group, catalyzing enzyme reactions, inducible cDNAs possibly involved in caffeoyl coenzyme A. < which lead to an inversion of the sugar the adap-tive process of light tolerance. configuration). Although we were able to modify and reduce the specific activi- ties of this en-zyme, changing one amino acid only, this was apparently not suffi- cient to alter position specificity or lea- ding to a significant change in substrate
Cladogram illustrating the distribution of selected glucosyltransferases involved in betacyanin biosyn- thesis.
64 65 Research Group: Biochemistry of Betalains (until 2001) Head: Willibald Schliemann
Group Members Betalains (red-violet betacyanins and yellow betaxanthins) are chromoalkalo- content increased in parallel with the In studies on vacuolar transport of beta- Naoko Kobayashi ids of chemotaxonomical importance. They functionally replace the anthocy- fresh weight, the cyclo-dopa 5-O-glu- xanthins, the inhibition pattern of the (PhD student until December 2001) anins in members of most families of the Caryophyllales. Betacyanins are also coside did not accumulate. The low MgATP-stimulated vacuolar uptake of Barbara Kolbe of commercial interest as food colorants. The main objective of our research amount of cyclo-dopa 5-O-glucoside the beet-specific miraxanthin V and vul- (technician) is the unravelling of betalain biosynthesis. After the characterization of the found originates from betanin, which is gaxanthin I (by 1 mM vanadate) and of Shiming Liu in equilibrium with cyclo-dopa 5-O- the unnatural (R)-phe- (guest scientist until December 2001) bifunctional tyrosinase and spontaneously proceeding steps, experiments to glucoside and betalamic acid under nylalanine-betaxan- detect the elusive dopa 4,5-dioxygenase are of particular interest as this enzy- slightly acidic conditions. This re sult is thin (by 0.1 µM bafilo- Collaborators me forms the chromophore betalamic acid, the key intermediate in betalain in contrast to previous data from mycin A1 and 5 mM biosynthesis. Hartmut Böhm Wyler et al. [Helv. Chim. Acta 67, 1348- NH4Cl) suggests the German Institute of Human Nutrition, Bergholz- 1355 (1984)], but in accordance with par ticipation of an Rehbrücke, Germany recent studies (Thomas Vogt, IPB) that ABC-like directly-ener - Yizhong Cai, Harold Corke A definite proof of the detection of dopa treatment with 30 µM methyl jasmonate. glucosyltransferases from red beets gized transport mecha - The University of Hong Kong, Hong Kong, People's Republic of China 4,5-dioxygenase activity in plants has not However, feeding of betalamic acid to accept betanidin, but not cyclo-dopa as nism and H+/antiport Inna Kuzovkina be achieved. In various enzyme assays the induced cells did not lead to the for- substrate. system, respectively. Timiryasev Institute of Plant Physiology, Russian Academy 14 of Sciences, Moscow, Russia with [ C]dopa, no soluble betalamic acid mation of yellow miraxanthin V, the Both systems have Enrico Martinoia, Markus Klein could be detected. However, radioacti- essential prerequisite for the detection The structures of betalains occurring been described in lite- Université de Neuchâtel, Switzerland vity was released from the assay proteins of a transient expression of a dopa di- in inflorescences of two Celosia varie- rature for the vacuo- Jürgen Schmidt, Thomas Degenkolb by alkaline hydrolysis; and by the addition oxygenase cDNA. ties (Celosia argentea var. cristata and lar uptake of endoge- Institute of Plant Biochemistry (IPB), Halle, Germany of (S)-Phe, labeled (S)-Phe-betaxanthin Celosia argentea var. plumosa) were nous luteolin glucuro- Victor Wray, Manfred Nimtz could be identified. This proved the for- To answer the question whether beta- elucidated in cooperation with part- nides in rye and flavo- German Research Centre for Biotechnology, Braunschweig, Germany mation of betalamic acid, but in very low nin biosynthesis in red beets proceeds ners of the IPB and from China. Three noid glucosides in bar - amounts. For a molecular approach to exclusively via cyclo-dopa or via cyclo- yellow pigments were found to be ley, respectively. The identify dopa dioxygenase using particle dopa 5-O-glucoside, the contents of immonium conjugates of betalamic research on betalains bombardment, a cell suspension culture be tanin and cyclo-dopa 5-O-glucoside acid with dopamine, 3-me th oxy - was terminated with of Tinospora cordifolia (Menisperma - in red beet hypocotyls were moni- tyramine and (S)-tryptophan. the end of 2001. < ceae) was selected which showed a 4- tored during eight weeks of plant fold in crease in dopamine content after development. Where as the betanin
Time course of betanin and cyclo-dopa 5-O-glucoside accumulation during the Flowering Celosia argentea var. cristata (above), structures of two new end- development of red beets. ogenous betaxanthins and Celosia argentea var. plumosa (below).
66 67 Research Group: Hydroxycinnamic Acids Head: Dieter Strack
Group Members Higher plants accumulate a wealth of hydroxycinnamate (HCA) conjugates, thesis and a physiological one trying to bidopsis. Homozygous transgenic lines Alfred Baumert mostly esters and amides. They are of prime ecological importance for plant divert one of the sinapine precursors, are developed that will be used for quan- (research scientist) survival. They protect plants against DNA-damaging UV light. Acylation of choline, into a new metabolic sink. The tification of sinapine and 1-sinapoylglu- Claus Lehfeldt anthocyanin pigments with HCAs results in an (intramolecular) copigmenta- two enzymes in focus are the SGT and cose in seeds. (PhD student until June 2001) tion effect, protecting these pigments against degradation. Soluble and cell the SCT. A cDNA encoding SGT was Carsten Milkowski isolated from cDNA libraries construc- As the SMT, the SCT belongs to the (postdoctoral position) wall-bound HCAs participate in plant defense against microbial attack. With ted from immature seeds and young group of SCPL enzymes. By a "homolo- Juliane Mittasch regard to their biosynthesis, HCAs are usually activated as coen-zyme A (PhD student since December 2002) seed lings of rape. The deduced SGT gy based cloning strategy", a full-length (CoA) thioesters or 1-O-acylglucosides (b-acetal esters), being the substrates Lilian Nehlin amino acid sequence indicated that SGT cDNA could be isolated from rape (guest scientist until June 2002) of the HCA transferases involved in formation of various conjugates. Our belongs to a distinct subgroup of gluco- seeds sharing about 85 % identity with Ingrid Otschik group is interested in structural and functional characterization of the UDP- syltransferases that catalyze the for- the SCT-cDNA from Arabidopsis. After (technician) glucose- and acylglucose-dependent glucosyl- and HCA transferases. mation of 1-O-acylglucosides. The SGT- expression in E. coli, the recombinant Diana Schmidt cDNA from rape was cloned and func- protein was shown to be in the inso- (PhD student since August 2001) tionally expressed in E. coli. The recom- luble fraction.As this is one of the main binant SGT carrying the His-tag at the C- problems with the class of SCPL pro- Collaborators Cloning of the 1-sinapoylglucose:mala- secondary metabolic pathways in plants. terminus was purified. The enzyme sho- teins, the optimization of heterologous te sinapoyltransferase (SMT) gene from wed a molecular mass of 60 kDa (gel fil- ex pression has come into the focus of Diana Bowles Department of Biology, University of York, UK Arabidopsis thaliana and immunolocaliza- In an approach to immunolocalize the tration) and 62 kDa (electrophoresis), our present work. Clint Chapple tion of the SMT protein SMT protein, rabbit polyclonal anti- respectively. It exhibited a broad sub- Purdue University, West Lafayette, USA SMT catalyzes the formation of sinapoyl- bodies were raised against the recom- strate specificity, accepting cinnamate, In the physiological approach, bacterial Martin Frauen, Gunhild Leckband malate, one of the major phenylpro- binant SMT expressed in E. coli from the 4-coumarate, caffeate, ferulate and sina- genes (betA and betB) encoding choli- Nordeutsche Pflanzenzucht, Hans Georg Lembke KG (breeder), Hohenlieth, Germany panoid secondary metabolites accumu- corresponding Arabidopsis cDNA. Im - pate. DNA cassettes for the dsRNAi- ne oxidase have been introduced in Ernst Heinz lated by some members of the Brassi - muno blot analysis of proteins from diffe- mediated seed-specific suppression of Ara bidopsis and rape. It is assumed University of Hamburg, Germany caceae, e. g. Arabidopsis thaliana, rape rent Arabidopsis tissues showed that the SGT were constructed and cloned into a that the glycine betaine pathway will Knut Meyer, Paul V. Viitanen (Bras sica napus) or red radish (Raphanus SMT is produced in all plant organs, ex - binary vector. Plant transformation was compete for choline as substrate in DuPont Central Research and Development, Biochemical sativus). In cooperation with Clint Chap - cept in the seeds and young seedlings. performed by collaborators of the sina-pine synthesis. Choline feeding Intracellular localization of SMT within Ara bidopsis Sciences and Engineering, Wilmington, Delaware, USA rosette leaves. Cross sections were immunodeco- ple, we identified previously an Ara bi - Immunofluorescent labeling of Arabi dop - University of Goettingen (Ch. Moellers). experiments using im mature Christian Möllers rated with polyclonal monospecific antibodies rai- dopsis mutant, sng1 (sinapoylglucose sis leaf sections localized SMT to the Arabidopsis and rape embryos revea- sed against the recombinant SMT protein followed University of Göttingen, Germany by fluorescence labelled secondary antibody. A José Orsini accumulator 1), which is defective in syn- cen tral vacuoles of mesophyll and epi- As a result of sequence comparison ana- led that the level of free choline is limi- green fluorescent label within the vacuoles of Saaten Union Resistenzlabor GmbH, Leopoldshöhe, thesis of sinapoylmalate. We have cloned dermal cells (see figure). In accor-dance lyses, four homologous genes encoding ted. Thus, to provide choline in a non- mesophyll cells of wild-type leaves (A) is indicative Germany of the SMT protein (arrows). In contrast, in the the corresponding gene and have found with characteristics of SCPL proteins, we hydroxycinnamate glucosyltransferases limiting concentration for glycine Jürgen Schmidt, Sabine Rosahl vacuoles of mesophyll cells of the deletion mutant that it encodes a serine carboxypepti- conclude that Arabidopsis SMT is syn- were cloned from Arabidopsis. These betaine synthesis, we will suppress sng1 (B), defective in synthesis of sinapoylmalate, Institute of Plant Biochemistry, Halle, Germany the fluorescent signals are absent (bar = 50 µm). Joachim Schröder dase-like (SCPL) protein. Expression of thesized as a precursor protein that is genes were functionally expressed in E. SGT activity in transgenic plants ex - University of Freiburg, Germany SNG1 in E. coli demonstrated that it targeted to the endoplasmic reticulum. coli. According to the acceptor specifi- pressing betA and betB. This strategy Milton T. Stubbs encodes the SMT. This finding suggests The protein is probably glycosylated in city, we identified one of them (AtSGT1) will hopefully not only im prove rape- University of Halle, Germany that SCPL proteins have acquired novel the Golgi apparatus from where it is sub- with high affinity to sinapate, whereas seed used as healthy food but will en - Victor Wray functions in plant metabolism and pro- sequently routed to the vacuole. the remaining three displayed broader hance by the accumulated oxidation German Research Centre for Biotechnology, Braunschweig, Germany vides an insight into the evolution of acceptor specificity. Based on the cDNA product of choline, glycinebetain, the Cloning of the cDNAs encoding UDP-glu- sequence of AtSGT1, DNA cassettes for tolerance of rape to environmental cose:sinapate glucosyltransferase (SGT) the dsRNAi-mediated suppression of the stresses, such as salt, low temperature and 1-sinapoylglu-cose:choline sinapoyl- SGT, using a seed-specific (napine) and a or drought that often affect seed ger- transferase (SCT) from Arabidopsis thali- constitutive promoter (CaMV 35S), were mination and plant productivity. < ana and Bras si ca napus constructed. Both suppression con- This work is part of the BMBF project structs were used to transform Ara - "NAPUS 2000 - healthy food from trans- genic rape" and focuses on reduction of the antinutritive sinapine (sinapoylcholi- ne) content in rapeseed. We are fol- lowing two strategies. A molecular ap - proach (dsRNAi) aims to suppress the pivotal enzymatic steps of sinapine syn-
68 69 Publications, Books and Bookchapters, In press, Patents, Doctoral Theses, Diploma Theses
Publications (2001). co and tomato. Phytochemistry 54, 473-479 (2000). Biosynthese pflanzlicher Farbstoffe. Angew. Chem. Anwohnern der Vulkane unseres Planeten, Shaker Books and Book chapters in press Bachmann, A., Hause, B., Maucher, H., Garbe, E., Hause, B., Maier, W., Miersch, O., Kramell, R. & Maucher, H., Hause, B., Feussner, I. & Wasternack, C. 113, 3907-3911 (2001). Verlag Aachen (2000). Stenzel, I., Hause, B., Feussner, I. & Wasternack, C. Weichert, H., Wasternack, C. & Feussner, I. Strack, D. Induction of jasmonate biosynthesis in The allene oxide synthase of barley (Hordeum vul- Van Damme, E. J. M., Hause, B., Hu, J., Barre, A., Fester, T., Peerenboom, E., Weiss, M. & Strack, D. Transcriptional activation of jasmonate biosynthesis Jasmonate-induced lipid peroxidation in barley lea- arbuscular mycorrhizal barley roots. Plant Physiol. gare cv. Salome) leaves is developmentally regula- Rougé, P., Proost, P. & Peumans, W. J. Two distinct Multimedia-Präsentation Mycorrhiza, (2001). enzymes is not reflected at protein level. In: ves initiated by distinct 13-LOX forms of the chlo- 130, 1213-1220 (2002). ted. Plant J. 21, 199-213 (2000). jacalin-related lectins with a different specificity Advanced Research on Plant Lipids (Murata, N., ed.) roplast. Biol. Chem. 383, 1645-1657 (2002). and sub-cellular location are major vegetative sto- Strack, D. Enzymes involved in hydroxycinnamate Kluwer Academic Publishers, Dordrecht, 2002. Hause, B., Meyer, K., Viitanen, P. V., Chapple, C. & Mikkat, S., Milkowski, C. & Hagemann, M. The gene rage proteins in the bark of the mulberry (Morus metabolism. In: Methods in Enzymology, Vol. 335, Back, K., Jang, S. M., Lee, B.-C., Schmidt, A., Strack, D. Strack, D. Immunolocalization of 1-O-sinapoylglu- sll0273 of the cyanobacterium Synechocystis sp. nigra) tree. Plant Physiol. 130, 757-769 (2002). Flavonoids and Other Polyphenols (Packer, L., ed.) Stumpe, M., Stenzel, I., Weichert, H., Hause, B. & & Kim, K.-M. Cloning and characterization of a cose:malate sinapoyltransferase in Arabidopsis tha- strain PCC6803 encodes a protein essential for Academic Press, Sheffield, UK, pp. 70-81 (2001). Feussner, I. The lipoxygenase pathway in mycorrhi- hydroxycinnamoyl-CoA:tyramine N-(hydroxycin- liana. Planta 215, 26-32 (2002). growth at low Na+IK+ ratios. Plant Cell Environ. 23, Van Damme, E. J. M., Hu, J., Barre, A., Hause, B., zal roots of Medicago truncatula. In: Advanced namoyl) transferase induced in response to UV-C 549-559 (2000). Baggerman, G., Rougé, P. & Peumanns, W. J. Varma, A. K., Singh, A., Sudha, Sahay, N. S., Sharma, J., Research on Plant Lipids (Murata, N., ed.) Kluwer and wounding from Capsicum annuum. Plant Cell Hause, B., Stenzel, I., Miersch, O., Maucher, H., Purification, characterization, immunolocalization Roy, A., Kumari, M., Rana, D., Thakran, S., Deka, D., Academic Publishers, Dordrecht, 2002. Physiol. 42, 475-481 (2001). Kramell, R., Ziegler, J. & Wasternack, C. Tissue-speci- Milkowski, C., Baumert, A. & Strack, D. Cloning and and structural analysis of the abundant cytoplasmic Bharti, K., Hurek, T., Blechert, O., Rexer, K.-H., Kost, fic oxylipin signature of tomato flowers: allene oxide expression of a rape cDNA encoding UDP-gluco- beta-amylase from Calystegia sepium (hedge bind- G., Hahn, A., Maier, W., Walter, M., Strack, D. & Thorson, J. & Vogt, T. Glycosylated natural products. Baumert, A., Mock, H.-P., Schmidt, J., Herbers, K., cyclase is highly expressed in distinct flower organs se:sinapate glucosyltransferase. Planta 211, 883-886 weed) rhizomes. Eur. J. Biochem. 268, 6263-6273 Kranner, I. Piriformospora indica - an axenically cul- In: Carbohydrate based Drug Discovery (Wong, C.- Sonnewald, U. & Strack, D. Patterns of phenylpropan- and vascular bundles. Plant J. 24, 113-126 (2000). (2000). (2001). turable mycorrhiza-like endosymbiotic fungus. In: H., ed.). oids in non-inoculated and potato virus Y-inoculated The Mycota, IX, Fungal Associations (Hock, B., ed.), leaves of transgenic tobacco plants expressing yeast- Hause, B., Weichert, H., Höhne, M., Kindl, H. & Milkowski, C., Baumert, A. & Strack, D. Identification Vierheilig, H., Gagnon, H., Strack, D. & Maier, W. Springer-Verlag, Wien New York, pp. 125-150 (2001). derived invertase. Phytochemistry 56, 535-541 Feussner, I. Expression of cucumber lipid-body lipo- of four Arabidopsis genes encoding hydroxycinna- Accumulation of cyclohexenone derivatives in bar- Patents (2001). xygenase in transgenic tobacco: lipid-body lipoxy- mate glucosyltransferases. FEBS-Lett. 486,183-184 ley, wheat and maize roots in response to inocula- Vogt, T. Glycosyltransferases involved in plant Hause, B., Bessler, K., Kogel, K. & Wasternack., C. genase is correctly targeted to seed lipid bodies. (2000). tion with different arbuscular mycorrhizal fungi. secondary metabolism. In: Evolution of Metabolic Method of screening for agrochemicals. European Binarová, P., Cenklová, V., Hause, B., Kubátová, E., Planta 210, 708-714 (2000). Mycorrhiza 9, 291-293 (2000). Pathways. Recent Advances in Phytochemistry, Vol. patent 981245251. Lysák, M., Dolezel, J., Bögre, L. & Dráber, P. Nuclear Milkowski, C., Krampe, S., Weirich, J., Hasse, V., 34 (Romeo, J. T., Ibrahim, R., Varin, L., de Luca, V., g-tubulin during acentriolar plant mitosis. Plant Cell Ibdah, M., Krins, A., Seidlitz, H., Heller, W., Strack, D. Boles, E. & Breunig. K. D. Feedback regulation of glu- Vierheilig, H., Maier, W., Wyss, U., Samson, J., Strack, eds.) Elsevier Science, New-York, pp. 317-347 Milkowski, C., Baumert, A. & Strack, D. Verfahren zur 12, 433-442 (2000). & Vogt, T. Spectral dependence of flavonol and beta- cose transporter gene transcription in D. & Piché, Y. Cyclohexenone derivative- and phos- (2000). Beeinflussung des Sinapingehaltes in transgenen cyanin accumulation in Mesembryanthemum cry- Kluyveromyces lactis by glucose uptake. J. phate- levels in split-root systems and their role in Pflanzenzellen und Pflanzen. German patent Breunig, K. D., Bolotin-Fukuhara, M., Bianchi, M. M., stallinum under enhanced UV radiation. Plant Cell Bacteriol. 183, 5223-5229 (2001). the systemic suppression of mycorrhization in pre- 10034320.1 (2000). Bourgarel, D., Falcone, C., Ferrero, I., Frontali, L., Environ. 25, 1145-1154 (2002). colonized barley plants. J. Plant Physiol. 157, 593- Publications in press Goffrini, P., Krijger, J. J., Mazzoni, C., Milkowski, C., Nehlin, L., Möllers, C., Bergmann, P. & Glimelius, K. 599 (2000). Cacace, S., Schröder, G., Wehinger, E., Strack, D., Rosahl, S., Scheel, D., Schmidt, A. & Strack, D. Steensma, H. Y., Wesolowski-Louvel, M. & Zeeman A. Irmler, S., Schröder, G., St-Pierre, B., Crouch, N. P., Transient beta-gus and gfp gene expression and via- Schmidt, J. & Schröder, J. A flavonol O-methyltrans- Transgene Pflanzen mit erhöhter Resistenz gegen M. Regulation of primary matabolism in Hotze, M., Schmidt, J., Strack, D., Matern, U. & bility analysis of microprojectile bombarded micro- Vogt, T. Substrate specificity and sequence analysis ferase from Catharanthus roseus performing two Befall durch Phytopathogene. German patent Kluyveromyces lactis. Enzyme Microb. Technol. 26, Schröder, J. Indole alkaloid biosynthesis in spores of Brassica napus L. J. Plant Physiol. 156, define a polyphyletic origin of betanidin 5- and 6- sequential methylations. Phytochemistry 62, 127- 19846001.5 (2000). 771-780 (2000). Catharanthus roseus: new enzyme activities and 175-183 (2000). O-glucosyltransferase from Dorotheanthus bellidi- 138 (2003). identification of cytochrome P450 CYP72A1 as formis. Planta 214, 492-495 (2002). Ziegler, J., Stenzel, I., Hause, B. & Wasternack, C. Cai, Y., Sun, M., Schliemann, W. & Corke, D. Chemical secologanin synthase. Plant J. 24, 797-804 (2000). Pauk, J., Puolomatka, M., Tóth, K. L. & Monostori, T. Eckermann, C., Schröder, G., Eckermann, S., Strack, Allenoxidcyclase-Gen und dessen Verwendung zum stability and colorant properties of betaxanthin pig- In vitro androgenesis of triticale in isolated micro- Vogt, T. & Jones, P. Glycosyltransferases in plant natu- D., Schmidt, J., Schneider, B. & Schröder, J. Herstellen von Jasmonsäure. German patent ments from Celosia argentea. J. Agric. Food Chem. Jones, P. & Vogt, T. Glycosyltransferases in seconda- spore culture. Plant Cell Tiss. Org. 61, 221-229 ral products synthesis: characterization of a super- Stilbenecarboxylate biosynthesis: a new function in 10004468.9 (2000). 49, 4429-4435 (2001). ry plant product metabolism: tranquilizers and sti- (2000). gene family. Trends Plant Sci. 5, 380-386 (2000). the family of chalcone synthase-related proteins. mulant controllers. Planta 213, 164-174 (2001). Phytochemistry 62, 271-286 (2003). Chen, Y., Peumans, W. J., Hause, B., Bras, J., Kumar, Peumans, W. J., Hause, B. & Van Damme, E. J. M. The Walter, M. H., Fester, T. & Strack, D. Arbuscular Doctoral Theses M., Proost, P., Barre, A., Rougé, P. & Van Damme, E. J. Kobayashi, N., Schmidt, J., Nimtz, M., Wray, V. & galactose-binding and mannose-binding jacalin-rela- mycorrhizal fungi induce the non-mevalonate Krajinski, F., Hause, B., Gianinazzi-Pearson, V. & Ibdah, Mwafaq: Lichtinduzierte Flavonoid- und M. Jasmonic acid methyl ester induces the synthesis Schliemann, W. Betalains from Christmas cactus. ted lectins are located in different sub-cellular methylerythritol phosphate pathway of isoprenoid Franken, P. Mtha1, an arbuscule cell-specific plasma Betacyanakkumulation in Mesembryanthemum cry- of a cyto plasmic/nuclear chito-oligosaccharide bin- Phytochemistry 54, 419-426 (2000). compartments. FEBS-Lett. 477, 186-192 (2000). biosynthesis correlated with accumulation of the membrane H+-ATPase gene from Medicago trunca- stallinum. University of Halle-Wittenberg, ding lectin in tobacco leaves. FASEB J. 16, 905-907 yellow pigment and other apocarotenoids. Plant J. tula. Plant Biol. Department of Pharmacy, 15/5/2002. (U225-251) (2002). Kobayashi, N., Schmidt, J., Wray, V. & Schliemann, W. Riemann, D., Rontsch, J., Hause, B., Langner, J. & 21, 571-578 (2000). Formation and occurrence of dopamine-derived Kehlen, A. Cell-cell contact between lymphocytes Opitz, S., Schnitzler, J.-P., Hause, B. & Schneider, B. Kobayashi, Naoko: Contributions to betalain bio- Ezcurra, I., Wycliffe, P., Nehlin, L., Ellerstrom, M. & betacyanins. Phytochemistry 56, 429-436 (2001). and fibroblast-like synoviocytes induces lymphocy- Walter, M. H., Hans, J. & Strack, D. Two distantly Histochemical analysis of phenylphenalenone-rela- chemistry. New structures, condensation reactions, Rask, L. Transactivation of the Brassica napus napin tic expression of aminopeptidase N/CD13 and related genes encoding 1-deoxy-D-xylulose 5- ted compounds in Xiphidium caeruleum and vacuolar transport. University of Halle- promoter by AB13 requires interaction of the con- Landtag, J., Baumert, A., Degenkolb, T., Schmidt, J., results in lymphocytic activation. Adv. Exp. Med. phosphate synthases: differential regulation in (Haemodoraceae). Planta. Wittenberg, Department Biochemistry/Biotech - served B2 and B3 domains of AB13 with different Wray, V., Scheel, D., Strack, D. & Rosahl, S. Biol. 477, 57-66 (2000). shoots and apocarotenoid-accumulating mycorrhi- nology, 20/11/2002. cis-elements: B2 mediates activation through an Accumulation of tyrosol glucoside in transgenic zal roots. Plant. J. 31, 243-254 (2002). Peng, Z. F., Strack, D., Baumert, A., Subramaniam, R., ABRE, whereas B3 interacts with an RY/G-box. potato plants expressing a parsley tyrosine decar- Roitsch, T., Ehneß, R., Goetz, M., Hause, B., Hofmann, Goh, N. K., Chia, T. F., Tan, S. N. & Chia, L. S. Lehfeldt, Claus-Ulrich: Das Gen der Plant J. 24, 57-66 (2000). boxylase. Phytochemistry 60, 683-689 (2002). M. & Sinha, A. K. Regulation and function of extra- Wasternack, C. & Hause, B. Jasmonate - Signale zur Antioxidant flavonoids from leaves of Polygonum Sinapoylglucose: L-Malat-Sinapoyltransferase von cellular invertase from higher plants in relation to Stressabwehr und Entwicklung in Pflanzen. Biologie hydropiper L. Phytochemistry 62, 16-21 (2003). Arabidopsis thaliana (L.) Heynh. Fester, T., Hause, B., Schmidt, D., Halfmann, K., Lee, Y. K., Hippe-Sanwald, S., Jung, H. W., Hong, J. K., assimilate partitioning, stress responses and sugar in unserer Zeit 30, 312-320 (2000). (Ackerschmalwand): Klonierung durch T-DNA- Schmidt, J., Wray, V., Hause, G. & Strack, D. Hause, B. & Hwang, B. K. In situ localization of chi- signalling. Aust. J. Plant Physiol. 27, 815-825 (2000). Proels, R.K., Hause, B. & Roitsch, T. Novel mode of Tagging und Versuche zur Expression in Escherichia Occurrence and localization of apocarotenoids in tinase mRNA and protein in compatible and Wasternack, C. & Hause, B. Jasmonates and octade- hormone induction of tandem tomato invertase coli. University of Halle-Wittenberg, Department of arbuscular mycorrhizal plant roots. Plant Cell incompatible interactions of pepper stems with Schliemann, W., Cai, Y., Degenkolb, T., Schmidt, J. & canoids - signals in plant stress response and deve- genes in floral tissues. Plant Mol. Biol. Bio chemistry/Bio tech nology, 11/4/2001. < Physiol. 43, 256-265 (2002). Phytophthora capsici. Physiol. Mol. Plant Pathol. 57, Corke, H. Betalains of Celosia argentea. lopment. Progr. Nucleic Acid Research 72, 165- 111-121 (2000). Phytochemistry 58, 159-165 (2001). 221 (2002). Stenzel, I., Hause, B., Maucher, H., Pitzschke, A., Fester, T., Kiess, M. & Strack, D. A mycorrhiza- Miersch, O., Kramell, R., Ziegler, J., Ryan C.A. & responsive protein in wheat roots. Mykorrhiza 12, Lehfeldt, C., Amber, M. S., Meyer, K., Ruegger, M., Stephan, M., Bangerth, F. & Schneider, G. Transport Yamamoto, K.-I., Kobayashi, N., Yoshitama, K., Wasternack, C. Allene oxide cyclase transgenes 219-222 (2002). Cusumano, J. C., Viitanen, P. V., Strack, D. & Chapple, and metabolism of exogenously applied gibberellins Teramoto, S. & Komamine, A. Isolation and purifica- potentiate jasmonate biosynthesis and the wound- C. Cloning of the SNG1 gene of Arabidopsis reve- to Malus domestica Borkh. cv. Jonagold. Plant tion of tyrosine hydroxylase from callus cultures of response of tomato leaves. Plant J. 33, 577-589 Fester, T., Schmidt, D., Lohse, S., Walter, M. H., Giuliano, als a role for a serine carboxypeptidase-like pro- Growth Regul. 33, 77-85 (2001). Portulaca grandiflora. Plant Cell Physiol. 42, 969- (2003). G., Bramley, P. M., Fraser, P. D., Hause, B. & Strack, D. tein as an acyltransferase in secondary metabolism. 975 (2001). Stimulation of carotenoid metabolism in arbuscular Plant Cell 12, 1295-1306 (2000). Strack, D., Fester, T., Hause, B. & Walter, M. H. Eine Stenzel, I., Hause, B., Miersch, O., Kurz, T., Maucher, mycorrhizal roots. Planta 216, 148-154 (2002). unterirdische Lebensgemeinschaft: Die arbuskuläre Ziegler, J., Stenzel, I., Hause, B., Maucher, H., H., Weichert, H., Ziegler, J., Feussner, I. & Lehmann, K., Hause, B., Altmann, D. & Köck, M. Mykorrhiza. Biologie in unserer Zeit 31, 286-295 Hamberg, M., Grimm, R., Ganal, M. & Wasternack, Wasternack, C. Jasmonate biosynthesis and the Fester, T., Strack, D. & Hause, B. Reorganization of Tomato ribonuclease LX with the functional ER (2001). C. Molecular cloning of allene oxide syclase: The allene oxide cyclase family of Arabidopsis thaliana. tobacco root plastids during arbuscule develop- retention motif HDEF is expressed during pro- enzyme establishing the stereochemistry of octa- Plant Mol. Biol. ment. Planta 213, 864-868 (2001). grammed cell death processes including xylem dif- Strack, D. & Schliemann, W. Bifunctional polyphenol decanoids and jasmonates. J. Biol. Chem. 275, ferentiation, germination and senescence. Plant oxidases: novel functions in plant pigment biosyn- 19132-19138 (2000). Strack, D., Vogt, T. & Schliemann, W. Recent advances Hao, Q., Van Damme, J. M., Hause, B., Barre, A., Physiol. 127, 436-449 (2001). thesis. Angew. Chem. Int. Ed. 40, 3791-3794 (2001). in betalain research. Phytochemistry 62, 247-269 Chen, Y., Rougé, P. & Peumans, W. J. Iris bulbs express (2003). type 1 and type 2 ribosome-inactivating proteins Maier, W., Schmidt, J., Nimtz, M., Wray, V. & Strack, D. Strack, D. & Schliemann, W. Bifunktionelle Books and Book chapters with unusual properties. Plant Physiol. 125, 866-876 Secondary products in mycorrhizal roots of tobac- Polyphenol oxidasen: neuartige Funktionen in der Fester, T. Leben aus dem Feuer? Eine Reise zu den
70 71 Department: Administration and Technical Services Head: Lothar Franzen Secretary: Heide Pietsch
he department Working Groups Tof Ad mi nis tra - tion and Tech nical Finance Ser vices re pre - Head: Barbara Wolf Astrid Ortloff (until August 2002) sents the cen - Gudrun Schildberg tral infra struc - Burgunde Seidl (since October 2002) tu ral unit within the Kerstin Wittenberg (since May 2002) institute. Ad mi n - Personnel istrative main focu- Head: Kerstin Balkenhohl ses are personnel, Alexandra Burwig legal, and financial Cindy Maksimo (since April 2002) mat ters. Main tasks Rita Stelzer of the central ser- Kathleen Weckerle vices are purcha- General Administration sing and account Head: Rosemarie Straßner ma nagement, and maintenance of the the gardeners take care of the expe- Alexandra Burwig scientific library and chemical store. rimental plant material. Cindy Maksimo (since April 2002) Rita Stelzer Also the gardeners represent an Kathleen Weckerle essential component of the central Since the reestablishment of the services. The Technical Ser vices deal institute in the year 1992, most main Trainees with the buildings and properties. buildings were re sto red completely. Antje Olschewski The technical co-workers in parti- The main focus of the construction Clemens Schinke cular take care of new construc- works concentrated on the labora- Library tions, the maintenance of the tory and technology areas, all of Head: Andrea Piskol exis ting buildings and laborato- which are now well equipped. Jessica Ackermann (Trainee) ries, and the technical and Antje Werner (Trainee) scientific equipment of the Beside the work the redeve lopment Graphics & Photography laboratories. of the existing buildings, new con- Head: Christine Kaufmann structions were undertaken in the Annett Kohlberg The scientific library of the last years. Currently, a new building Construction and Maintenance institute offers excellent is under construction that will Head: Matthias Böttcher (until December 2002) possibilities for literature- accomodate new highly sensitive Detlef Dieckmeyer based research. The libra- analytical instruments. In the near Carsten Koth (since January 2002) Michael Kräge ry has subscriptions to 83 future, additional greenhouse facili- Jörg Lemnitzer of international research ties and a central service building Klaus-Peter Schneider journals and stocks will be constructed. Catrin Timpel approximately 5.000 hardback Eberhard Warkus books. A reading hall with 18 in - Altogether, the institute of fers a Electronics ternet-connected computers and best possible infrastruc ture for its Holger Bartz five individual rooms are also diverse research projects. < Hans-Günter König avialable. Ronald Scheller Gardening In addition to an experimen- Head: Iris Rudisch tal field area, a series of fully Martina Allstädt air-conditioned green hou- Nicole Mühlwald (Trainee) ses and phytochambers are Christian Müller (since April 2002) Kristina Rejall (since June 2002) available for the re search Steffen Rudisch programs. In these areas Katja Scheming (Trainee since August 2002) Andrea Voigt (Trainee since August 2002)
72 73 Resources and Investments Staffing Schedule
Research grants listed on this and the following pages were given by in Mio. Euro in % Staffing schedule 2000 2001 2002 Total Average Number of members on annual average 166 167 169 502 167 AFNG Arabidopsis Functional Basic Financing Funds Genomics Network (DFG) Full-time employees in % 77 77 75 229 76 Personnel 13,0 32,4 BMBF Bundesministerium für Bil dung Part-time employees in % 23 23 25 71 24 und Forschung - Fede ral Consumables 6,2 15,5 Ministry of Education and Number of established posts 89 89 92 270 90 Research Grants / Subsidies 0,3 0,7 BML Bundesministerium für Ver - Temporary employees (budget) 19 11 20 50 17 braucher schutz, Ernährung und Investments 13,8 34,3 Landwirtschaft - Federal Employess remunerated by third parties subsidies Ministry of Consumer 42 44 38 124 41 “University Science Funds Programme” 1,2 3,0 (avarage) Protection, Food and Agricultur (HWP) Employees remunerated by "Universitiy Special Funds 1 - - 1 1 BPS BASF Plante Science GmBH Subtotal 34,5 Programme III" (Hochschulsonderprogramm III / HSP III)
D-B Foundation Gottlieb Daimler and Karl Benz Employees remunerated by "University Science Funds - 11 9 20 10 Foundation Funds from external sources Programme" (Hochschulwissenschaftsprogramm / HWP) DBU Deutsche Bundesstiftung BMBF 1,3 3,2 Proportion of female employess in % 60 59 61 180 60 Umwelt MK-LSA 0,8 2,0 Personnel fluctuation rate in % 16,4 10,4 13 39,8 13 DFG Deutsche Forschungsgemeinschaft DFG 2,4 6,0 Avarage age of employees 40 39 39 118 39 DAAD Deutscher Akademischer Austauschdienst - German Industry 0,6 1,5 Scholarship/fellowship holders 10 6 5 21 7 Academic Exchange Service EU 0,5 1,2 Vocational training Elsevier Elsevier Science Publisher other 0,1 0,2 commercial area 2 2 2 6 2 horticultural area 3 2 3 8 3 EU European Union Subtotal 5,7 14,1 library 1 2 2 5 2 Firmenich Total 40,2 100 Successfully completed vocational training 4 - - 4 1 GABI Genom Analyse im Biologischen System Pflanze Avarage number of apprentices 6 6 7 19 6
GTZ Gesellschaft für Technische Zusammenarbeit Investments Equipment 5,8 Hopsteiner Building 8,0 HSP III Hochschulsonderprogramm III Total 13,8 Humboldt Alexander von Humboldt Foundation Foundation HWP Hochschulwissenschaftsprogramm
Icon genetics
KWS KWS SAAT AG
MK-LSA Kultusministerium des Landes Sachsen-Anhalt - Ministry of Edu cation and Cultural Affairs of the State of Saxony Anhalt
PPP Projektbezogener Personenaustausch (DAAD)
Probiodrug Probiodrug AG
SFB 363 Sonderforschungsbereich 363 - Collaborative Research Centres
VW Foundation Volkswagen Foundation
74 75 Use of Funds from External Sources
Amount Personnel posts Amount Personnel posts Project & Head of Project Total duration Financed by 2000 - 2002 Project & Head of Project Total duration Financed by 2000 - 2002 (in Euro) financed (in Euro) financed
Chrom-(II)-mediated reactions 02/03 DAAD / PPP 5.700 0 Department of Natural Product Biotechnology (Prof. L. Wessjohann) Hungary Jasmonate biosynthesis regulation 99/04 DFG / SPP 111.000 1 Daimler Benz fellowship 2002 D-B Foundation 1.700 1 (Prof. C. Wasternack & O. Miersch) (Prof. L. Wessjohann)
Glutamate cyclase 01/03 Probiodrug 30.700 0 Subtotal: 1.000.800 8 (Prof. C. Wasternack)
Allenoxidcyclase 01/02 Firmenich 60.900 1 (Prof. C. Wasternack) Department of Stress and Developmental Biology Heavy-metal tolerance Papaver somniferum 00/01 DFG / SFB 363 57.300 1 02/04 DFG / SFB 363 96.400 1 (Prof. T. Kutchan) (D. Neumann & S. Clemens)
Papaver somniferum 01/02 DFG 117.900 1 Pathogen defense-related genes 98/00 DFG 30.000 1 (Prof. T. Kutchan) (Prof. D. Scheel)
Functional genomics 00/02 DFG 26.300 0 Signal transduction 02/04 DFG / SFB 363 201.100 1 (G. Herrmann) (Prof. D. Scheel)
Analysis of genes 00/02 Icon Genetics 230.900 1 Oxidative burst 99/00 DFG / 25.500 1 (Prof. T. Kutchan) (Prof. D. Scheel) Innovationskolleg
Molecular genetics of isoquinoline 01/04 DFG 67.600 2 Plant peptides 2000 DFG 460.200 1 alk.biosynth. (Prof. T. Kutchan) (Prof. D. Scheel)
Cellular signalling 02/04 DFG / MLU 24.100 1 Signal transduction 99/00 DFG 2.900 1 (Prof. T. Kutchan) (Prof. D. Scheel) Transformation and regeneration of Papaver 02/03 DFG 57.300 2 Chromatin and gene regulation 99/01 DFG / SFB 363 68.600 1 somniferum (S. Frick) (Prof. D. Scheel) Elicitor receptors Modulation of jasmonates by transgenic 02/04 DFG / SFB 363 175.500 1 99/01 DFG 54.100 2 plants (Prof. C. Wasternack & O. Miersch) (T. Nürnberger)
Salvia fragrances 02/03 DBU 13.850 1 CRISP 01/04 EU 154.400 1 (Prof. T. Kutchan) (Prof. D. Scheel)
Subtotal: 973.350 12 Heavy metal tolerance and silicon 00/04 MK-LSA 67.700 1 (U. zur Nieden)
Non-host resistance 98/02 KWS 120.300 1 Department of Bioorganic Chemistry (T. Nürnberger) Conformation of brassinosteroids 99/00 DFG 6400 1 The role of jasmonates in pathogene defen- 01/03 DFG 74.700 1 (A. Porzel, W. Brandt [MLU]) se (Prof. D. Scheel)
New bioactive natural products from endemically 97/02 DFG / GTZ 49.700 0 Jasmonate-insensitive mutant 99/03 MK-LSA 64.700 1 occuring Yemenian plants (J. Schmidt, G. Adam) (Prof. D. Scheel, S. Berger)
HEA(N)TOS 00/03 BMBF 264.700 2 Gene silencing 00/01 MK-LSA 91.400 1 (Prof. L. Wessjohann, Prof. G. Adam) (Prof. D. Scheel)
Structural elucidation and combinatorial 2000 MK-LSA / HWP 490.800 0 Arabidopsis halleri 00/03 DFG 60.300 1 chemistry (Prof. L. Wessjohann) (S. Clemens)
COMBIOCAT 01/04 EU 142.100 2 Ozone signaling 2001 DAAD / PPP 3000 0 (Prof. L. Wessjohann) (Prof. D. Scheel) Finnland
EPILA 01/03 EU 29.800 2 Metallophytes 01/03 EU 104.100 1 (W. Brandt) (S. Clemens)
Fungi excursion 2001 DFG 1.100 0 Biomineralisation 01/03 DFG 49.100 1 (N. Arnold) (D. Neumann)
MCR ligand synthesis 02/03 DAAD / Probral 8.800 0 Heat stress proteins 99/00 DFG 8.300 1 (Prof. L. Wessjohann) (D. Neumann)
76 77 Use of Funds from External Sources
Amount Personnel posts Amount Personnel posts Project & Head of Project Total duration Financed by 2000 - 2002 Project & Head of Project Total duration Financed by 2000 - 2002 (in Euro) financed (in Euro) financed
Signals, delivery and response 97/00 EU 68.600 2 Phytochemistry 02/04 Elsevier 8.000 1 (T. Nürnberger) (Prof. D. Strack)
Humboldt fellowship 01/02 Humboldt 5.600 0 Stable transformation of Medicago truncatu- 2002 MK-LSA 47.300 0 (Prof. D. Scheel) Foundation la (B. Hause)
Cooperation wtih South Africa 01/04 VW Foundation 40.000 0 Subtotal: 967.700 13 (T. Nürnberger)
NODO 02/04 EU 50.700 1 (S. Rosahl) Joint projects Receptor kinases 02/04 DFG / AFNG 47.600 2 Profiling of metabolites, proteins and peptides (T. Nürnberger) Dep. Stress and Developmental Biology and 00/04 BMBF / GABI 588.200 4 Dep. Bioorganic Chemistry Arabidopsis thaliana interactions 01/02 BPS 10.900 0 (Prof. D. Scheel) (S. Clemens)
Bioinformatics and Mass Spectronomy 02/07 BMBF 100.000 6 HUMULUS (Prof. D. Scheel) Dep. Bioorganic Chemistry and 01/02 Hopsteiner 9.900 0 Dep. Natural Product Biotechnology GABI-NONHOST 02/06 BMBF 109.700 4 (Prof. D. Scheel) (F. Stevens, Prof. L. Wessjohann & J. Page)
Pathogene defense in Arabidopsis thaliana 2002 MK-LSA 10.800 0 “Analytica 2000” 2000 MK-LSA 11.200 0 (S. Rosahl) (E. Peerenboom)
Subtotal: 2.180.700 34 “Achema 2000” 2000 MK-LSA 1.300 0 (E. Peerenboom) Department of Secondary Metabolism Public Understanding of Sciences and Donors Association Humanities (PUSH) - Multimedia project 2001 for the Promotion of 6.200 0 about the mycorrhiza Sciences and Huma - Betalains 99/01 DFG 61.800 1 (W. Schliemann & Prof. D. Strack) (T. Fester & E. Peerenboom) nities in Germany
Betanidin-Glucosyltransferases 01/03 DFG 121.900 2 Subtotal: 616.800 4 (T. Vogt)
TIMBER 98/00 EU 5.600 1 Projects granted, total: 5.739.350 71 (M. Walter)
Endomycorrhiza 98/01 DFG 24.000 1 (W. Maier & Prof. D. Strack) General view Metabolism of isoprenoids 00/04 DFG 90.500 1 BMBF 1.331.000 (M. Walter & T. Fester)
NAPUS 2000 99/04 BMBF 378.100 2 MK-LSA 785.200 (Prof. D. Strack)
Jasmonates in the development of barley 99/01 DFG 40.600 1 DFG 2.401.700 (B. Hause & Prof C. Wasternack)
The role of jasmonates during the establishment 00/04 DFG 83.500 1 Industry 573.300 of mycorrhiza (B. Hause & Prof. D. Strack)
Carotenoid biosynthesis in arbuscular 00/04 DFG 82.000 1 EU 555.300 mycorrhizal roots (T. Fester)
Metabolite profiling 02/04 DFG 24.400 1 Other sources 92.850 (W. Schliemann)
78 79 Guest Researchers and Fellows
Name Country Period Name Country Period Prof. Tamás Patony Hungary 13.10.2002 - 22.10.2002 Department of Natural Product Biotechnology (DAAD Fellow) Prof. Guillermina Abdala Argentina 08.06.2001 - 16.07.2001 Prof. Luay Rashan Jordan / Iraq 01.07.2002 - 31.08.2002 Dr. Maged Abou-Hashem Egypt 01.07.2002 - 22.10.2002 (Humboldt Fellow) Dr. Oscar Dorneles Rodriguez Brazil 01.04.2002 - 20.09.2002 Arysyak Abrahamian Armenia 04.07.2000 - 31.12.2000 (CAPES Fellow) (DAAD Fellow) Lars Seipold Germany 01.01.2002 - 31.05.2002 Nigel Bailey UK 19.11.2001 - 14.12.2001 Prof. Tran Van Sung Vietnam 01.07.2002 - 18.12.2002 Dr. Davide Berlanda Italy 05.06.2001 - 21.06.2001 Trin Thi Thuy Vietnam 20.11.2001 - 19.11.2002 Hubert Chassaigne France 15.01.2000 - 31.12.2000 (Humboldt Fellow) Larissa Vasilets Russia since 28.11.2002 Dr. Kum-Boo Choi Korea since 07.10.2002 (Humboldt Fellow) Dr. Svetlana Zakharova Russia 30.10.2002 - 31.12.2002 Predro Salvador de Rocha UK 10.04.2000 - 20.04.2000 (FEBS Fellow) Department of Stress and Developmental Biology Satinder Gitta Canada 05.04.2000 - 04.06.2000 Reetta Ahlfors Finland since 08.07.2002 (DAAD Fellow) Kristin Krukenberg USA 23.09.2002 - 15.07.2003 (Fulbright Fellow) Dr. Susanne Berger Germany 01.04.2001 - 31.03.2002 (DFG Fellow) Tamara Krupnova Kazakhstan 01.10.1999 - 30.04.2000 (DAAD Fellow) Anne-Claire Cazalé France 01.02.2000 - 31.12.2001 (Humboldt Fellow) Anan Onaroon Thailand 26.07.1999 - 30.09.2002 (DAAD Fellow) Clarice de Figueiredo Brazil 01.11.1999 - 30.09.2001 Matjaz Oven Slovenia 03.08.1999 - 31.07.2001 01.09.2001 - 30.09.2001 Anna Drobek Poland 26.02.2002 - 30.04.2002 Suppachai Samapitto Thailand 04.05.2000 - 03.05.2001 (DAAD Fellow) Dr. Emiko Harada Japan since 22.02.2002 (Humboldt Fellow) Anastasia Tkatcheva Canada 01.10.2001 - 28.02.2002 (SFB Fellow) Emma Jack Netherlands 12.03.2001 - 20.04.2001 Prof. Gülacti Topku Turkey 02.04.2000 - 02.07.2000 (DAAD Fellow) Dr. Anano Dinakar Karve India 16.11.1999 - 14.02.2000 Prof. Luc Varin Canada 01.10.2002 - 31.01.2003 (Humboldt Fellow) Dr. Magdalena Krzymowska Poland 01.08.1999 - 30.06.2002 Dr. Ana Vigliocco Argentina 01.04.2002 - 31.05.2002 (Humboldt Fellow) Dr. Bathany Zolman USA 15.08.2002 - 31.10.2002 Ma. Shaokang Singapore 06.05.2001 - 21.06.2001 (SFB Fellow) Srpryia Paranthaman India 25.10.2002 - 20.12.2002 Department of Bioorganic Chemistry (Humboldt Fellow) Prof. Antonio Luiz Braga Brazil 06.04.2002 - 21.04.2002 Lizelle Piater South Africa 01.06.2002 - 29.07.2002 (CAPES Fellow) Joe Chou Hung Sim Singapore 06.05.2001 - 21.06.2001 Tran Van Chien Vietnam since 07.10.2002 Claudia Simm Germany since 01.10.2000 Marco Aurelio Dessoy Brazil 01.02.2001 - 30.06.2002 (Fellow, Graduierten Kolleg) (DAAD Fellow) Anne Varet France 01.01.2002 - 30.04.2002 Csongor Hajdu Hungary 29.01.2001 - 31.07.2001 (Erasmus + DAAD Fellow) 19.08.2002 - 13.12.2002 Dubravko Jelic Croatia 10.03.2002 - 22.03.2002 Department of Secondary Metabolism Stijn Jan Freddy Desmyter Netherlands 10.01.2000 - 05.02.2000 Myint Myint Khine Myanmar (Burma) since 04.09.2002 (Daimler-Benz Fellow) Dr. Shiming Liu China 13.08.2001 - 13.06.2002 Lazlo Merczs Hungary 10.11.2002 - 11.12.2002 (DAAD Fellow) Dr. Nirmal Sahay India 10.01.2000 - 31.12.1999 Prof. Károly Micskei Hungary 17.06.2002 - 26.06.2002 (DAAD Fellow) Dr. Sudha Sahay India 24.09.1999 - 31.08.2001 Nguyen Hoang Anh Vietnam 01.09.2000 - 31.08.2001 Diana Schmidt (Fellow, Bio Service GmbH, EU and the State of Germany since 01.08.2001 Nguyen Hong Thi Van Vietnam since 17.04.2002 Saxony Anhalt)
80 81 Press and Public Relations Head: Sylvia Pieplow
Participation in trade shows exhibition pre-sented the history of Group members In 2000, the institute exhibited several plant breeding and the increasing role of Gesine Krüger pro jects at exhibitions and trade shows. gene technology for identifying and cre- (Head until März 2002) These activities were planned and orga- ating new kinds of productive and resi- Jana Krupik nized by Ellen Peerenboom. In March, stant plants. (Assistent and Webmaster since November 2000) the IPB participated in one of the biggest Ellen Peerenboom international conventions for biotechno- The exhibition “Life Science Art” in July (Head until Juli 2001) logy, “Bio 2000” in Boston. 2000 examined the theme of the human being that lies behind the scientific re - Together with the universities of searcher and his work. Silvia Stabel, Halle and Magdeburg and several painter and ex-scientist, displayed pain- other scientific institutes, our re - tings with scientific themes and objects searchers presented their work from the artist's perspective. The pain- at the Saxony Anhalt booth at tings' message about the beauty and aes- “Ana lytika” (in 2000 and thetic qualities of molecules, cells and 2002) in Munich and at scientific mo tifs was underlined by short “Achema” (in 2000) in explanations and quotations from re - Fran k furt. In addition to searchers. Many visitors were very the projects on display, impressed by the exhibits, which were Claus Wasternack chai- shown under titles like “Alphabet of life”, red a workshop to the “Hope” or “Orientation”. topic "Plant Bio tech - nology - Novel Food" Public events - a bridge to the peo- at a meeting held to - ple “Biotechnika” 2001 in Hannover gether with “Analytika As in the years before, the IPB participa- 2000”. ted in 2000 and 2001 in the “Science day” on the market square in Halle. Our Furthermore, in 2001 researchers presented the work of the the IPB participated institute by displaying posters and small- in “Biotechnika” in scale experiments. The event, organized Han nover, Germany's by the municipality and the University of most important inter- Halle, led to increased contact and national biotechnology discussion with interested citizens. exhibition. At all the ex hibitions and trade A similar presentation of scientific insti- shows, visiting scientists tutes on the market square in Halle was and journalists showed keen celebrated on the Uni versity’s 500th interest in the institute's Anniversary in June 2002. Research ers of work. As a result, several arti- the IPB displayed and explained living mo - cles were published in diffe- dels of mycorrhiza - a close partnership rent newspapers and journals. between plants and fungi. In addition, vi - sitors had the possibility to see mycor- Exhibitions at the frontier rhized plant cells under the microscope. between science and art A display of computer simulations about Hosting the exhibition “Gene world the characteristics and behavior of pro- and nutrition” in June 2000, presented teins was also shown. by the Alimentarium Food Museum of Vevey, Switzerland, was a great success In February 2000, members of the insti- for the IPB. More than 1000 guests visi- tute organized a charity concert for the ted the institute to view and critically community kitchen of St. Elizabeth's discuss the interactive exhibits. This Hospital in Halle. This community insti- “Science day” 2001 on the market square in Halle
82 83 tution provides about 70 poor people all secondary schools of Saxony Anhalt, Research, the Ministry of Educationand Scheel, director of the institute, spoke with a warm meal every day. As a result and afterwards as a result of numerous Cultural Affairs of Saxony Anhalt, the about the successful scientific tradition of this classical concert, the IPB collea- articles in the regional and national Leibniz Association, the city council of of the IPB. Three scientific reports and a gues proudly donated the money for 325 press, to many interested private citi- Halle and scientists from all over the concert by the chamber orchestra of the meals to the hospital. zens. This year an update and production world attended the official ceremony university completed the program. of an English version are planned. and expressed their best wishes for the After wards the institute members had a Under the motto "Green gene techno- future. In a ceremonial address, Dierk party together with all invited guests. < logy - prospects and risks", 24 teachers As in previous years, the IPB organized for chemistry and biology had the possi- many guided tours through the institute bility to learn more about new methods for school classes and senior groups in and molecular techniques of gene trans- 2000 to 2002. In addition, all of the four fer into plants in May 2002. The training, scientific departments sponsored several held under the auspices of the Central periods of practical training, which al- Marketing Organization of German Agri - lowed many high-school students to gain Publications Uni versity’s 500th Anniversary in June 2002 on the Benno Parthier celebrated his 70th birthday in market square in Halle cultural Industries, was organized by the insight into lab work and to try out Peerenboom, E. Staffellauf in der Pflan zenzelle. In: Berichte aus der August 2002 IPB. experiments on the bench. Wissenschaft, Deutscher Forschungsdienst, Bonn, pp. 13-15 (2000). Peerenboom, E. Zusatz für Farbindustrie bald aus Leinöl. WGL-Journal 1, Instead of the annual "Science day", in Since the beginning of 2001, the IPB has p. 25 (2000). 2002 the university and the other re- had a new corporate design. The inter- Peerenboom, E. & Stabel, S. Life Science Art. Leibniz 4, Sonderbeilage search institutes of Halle celebrated a new nally designed logo was successfully in - (2000). event - the "Long Night of Sciences". At troduced and promptly accepted by peo- Pieplow, S. Pflanzliche “Staubsauger” ziehen Schwermetalle aus dem this day in September, the institute's ple from both within and outside the Boden. Chemie.DE www.chemie.de/news/d/16725/ (2002). doors were open from 7 pm to midnight IPB. Since then, all of the letterheads on Scheel, D., Frohberg, K., Peerenboom, E. & Wakenhut, U. Traditionen to welcome more than 300 members of business letters, flyers, publicity brochu- verbunden mit neuen wissenschaftlichen Potentialen. In: the public. Visitors enthusiastically parti- res and business cards display the new Wirtschaftsstandort Halle, Europäischer Verlag, Darmstadt, pp. 92-97 cipated in guided tours through the labs logo. In addition, the institute's homepa- (2000). and greenhouses of the IPB. In addition, ge was completely reorganized and re- experiments were displayed in the foyer, newed. The new version in German and guests viewed the IPB's collection of co- English has been online since May 2001. Press releases l or ful cell cultures and had the possibi- Leckere Gene? Gen-Welten Ernährung Son derausstellung am IPB lity to learn how a confocal laser-scan- Celebrations (E. Peeren boom, 06.06.2000) ning microscope works. Be cause of the The year 2002 was a time of many cele- “Long Night of Sciences” 2002. Guests were verry Eine Symbiose aus Kunst und Wissen schaft: Sonder ausstel lung “Life Congratulations for Günter Adam. His 70th birth- interested in the interactive CD about the mycor- great success of this event, the "Long brations for the IPB. The institute hono- Science Art” am IPB (E. Peerenboom, 03.07.2000). day was celebrated in December 2002 rhiza. Night of Sciences" will be come an annual red two former members, each a cele- Prof. Dr. Ludger Wessjohann wird neuer Abteilungsleiter der event in Halle. brity due to his personal qualities and Abteilung Natur stoff chemie am Leibniz-Institut für Pflan zen bio chemie scien tific lifework, with a splendid collo- (E. Peerenboom, 26.10.2000). Public projects quium. In August, Benno Parthier, the PlantMetaNet - neues Forschungs netz werk - Vier führende The IPB participated in the competition institute's former director and president Institute auf dem Gebiet der Pflanzenforschung vereinbaren "Public Understanding of Sciences and of Germany's biggest and oldest acade- Kooperation (E. Peerenboom, 06.06.2001). Humanities" (PUSH) with a multimedia my, the German Academy of Natural Wissenschaftler entwickeln Lernmaterial für Schüler - Lern-CD für project about mycorrhizal symbiosis. Scien tists Leopoldina, celebrated his das Fach Bio logie: Mykorrhiza (G. Krüger, 12.12.2001). The interactive course about this fasci- 70th birthday together with his former IPB feiert sein 10jähriges Gründungs jubi läum (J. Krupik, 21.05.2002) nating biological interaction was produ- colleagues and the entire institute. ced by Thomas Fester and Ellen Peeren - Günter Adam, former head of the IPB ehrt langjährigen Direktor Prof. Dr. Benno Parthier (J. Krupik, 27.08.2002) boom and was designed for students and department Natural Product Chemistry, interested nonscientists alike. As one of also turned 70 in December. Lange Nacht der Wissenschaften “Blick ins Innere der Pflanze the 22 final winners selected, the project (S. Pieplow, 18.09.2002) was sponsored by the "Donors Asso - In May 2002, the institute celebrated its Festveranstaltung zu Ehren von Pro fes sor Adam (S. Pieplow, ciation for the Promotion of Sciences own birthday and foundation ten years 09.12.2002) and Human ities in Germany". Interactive ago on a large scale. Representatives of "Long Night of Sciences" 2002. Prof. Dierk Scheel Pflanzliche “Staubsauger” ziehen Schwer metalle aus dem Boden IPB’s 10th birthday in May 2002 guided the visitors through the institute. CD's were send away in 2001, at first to the Federal Ministry of Education and (S. Pieplow, 09.12.2002).
84 85 Map & Impressum
How to get to the IPB
Annual Report 2000 - 2002 of the Institute of Plant Biochem - istry Halle (Saale), July 2003
Publisher: Institute of Plant Biochemistry Weinberg 3 06120 Halle (Saale) Germany www.ipb-halle.de Editor: Sylvia Pieplow Press and Public Relations Phone: +49 (0) 3 45 - 55 82 11 10 Fax: +49 (0) 3 45 - 55 82 11 19 email: [email protected] [email protected]
Layout & Design: Jana Krupik Sylvia Pieplow Graphics & Pictures: Christine Kaufmann Annett Kohlberg Bettina Hause and others Copyright © 2003, all rights reserved Leibniz Institute of Plant Bio chemistry (IPB), Halle, Germany; No parts of this publication may be reproduced by any mechanical, photographic, electronic process, or in form of a photographic recording, nor may be stored in a retrieval system, transmitted or otherwise copied for public or private use, without written permission from the publisher.
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