NITROGEN FIXATION: FROM MOLECULES TO CROP …978-0-306-47615-0/1.pdf · Nitrogen Fixation: From Molecules to Crop Productivity Proceedings of the International Congress on Nitrogen

NITROGEN FIXATION: FROM MOLECULES TO CROP PRODUCTIVITY

Current Plant Science and Biotechnology in Agriculture

VOLUME 38

Scientific EditorR.J. Summerfield, The University of Reading, Department of Agriculture, P.O. Box 236, ReadingRG6 2AT, Berkshire, UK

Scientific Advisory BoardD.F. Bezdicek, Washington State University, Pullman, USAJ. Denecke, University of York, York, UKG.O. Edmeades, CIMMYT, MexicoJ. Hamblin, The University of Western Australia, Nedlands, WA, AustraliaH.-J. Jacobsen, Universität Hannover, Hannover, Germany

Aims and ScopeThe book series is intended for readers ranging from advanced students to senior research scientistsand corporate directors interested in acquiring in-depth, state-of-the-art knowledge about researchfindings and techniques related to all aspects of agricultural biotechnology. Although the previous vo-lumes in the series dealt with plant science and biotechnology, the aim is now to also include volumesdealing with animals science, food science and microbiology. While the subject matter will relatemore particularly to agricultural applications, timely topics in basic science and biotechnology willalso be explored. Some volumes will report progress in rapidly advancing disciplines through procee-dings of symposia and workshops while others will detail fundamental information of an enduringnature that will be referenced repeatedly.

Nitrogen Fixation:From Molecules toCrop ProductivityProceedings of the International Congress on Nitrogen Fixation,Foz do Iguaçu, Paraná, Brazil, September 12–17, 1999

Edited by

FABIO O. PEDROSADepartamento de Bioquímica,Universidade Federal do Paraná,Curitiba, Paraná, Brazil

MARIANGELA HUNGRIAEmbrapa Soja,Londrina, PR, Brazil

GEOFFREY YATESDepartamento de Bioquímica,Universidade Federal do Paraná,Curitiba, Paraná, Brazil

and

WILLIAM E. NEWTONDepartment of Biochemistry,Virginia Polytechnic Institute and State University,Blacksburg, Virginia, U.S.A.

KLUWER ACADEMIC PUBLISHERS NEW YORK, BOSTON, DORDRECHT, LONDON, MOSCOW

eBook ISBN: 0-306-47615-0Print ISBN: 0-7923-6233-0

©2002 Kluwer Academic PublishersNew York, Boston, Dordrecht, London, Moscow

Print ©2000 Kluwer Academic Publishers

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Dordrecht

INTERNATIONAL CONGRESSON NITROGEN FIXATION

was organized by

UNIVERSIDADE FEDERAL DO PARANÁ, BRAZIL

and supported by

The Brazilian Agriculture Research Company - EMBRAPAConselho Nacional de Desenvolvimento Científico e Tecnológico - CNPq

Financiadora de Estudos e Projetos - FINEPParaná Tecnologia

The Brazilian Society of Biochemistry and Molecular Biology - SBBqThe International Congress on Nitrogen Fixation, Paris, France

The Federal University of Paraná - UFPRThe Microbiological Resources Center - UNESCO/UNEP/ICRO

The International Foundation for Science (IFS)The Third World Academy of Sciences

The United States Department of Agriculture (USD A)Fundação da Universidade Federal do Paraná - FUNPAR

Liphatech Inc. (USA)Industria Biosoja de Inoculantes Ltda (Brazil)

PE BiosystemsAmersham - Pharmacia Biotech. do Brasil Ltda

Life Technologies do Brasil Ltda.Labtrade do Brasil Ltda

Financial support for the compilation and editing of this Book of Proceedings wasprovided by Financiadora de Projetos e Estudos (FINEP) and

by Paraná Tecnologia.

v

vi

INTERNATIONAL CONGRESSON NITROGEN FIXATION

was organized by the following committees

NATIONAL ORGANIZING COMMITTEE

Fábio O. Pedrosa (President, Universidade Federal do Paraná, Curitiba), Mariangela Hungria

(General Secretary, EMBRAPA Soja), Elaine M. Benelli (UFPR, Curitiba), Leda S. Chubatsu

(UFPR, Curitiba), Hidevaldo B. Machado (UFPR, Curitiba), Cláudio Rigo (UFPR, Curitiba),

Liu U. Rigo (UFPR, Curitiba), Emanuel M. Souza (UFPR, Curitiba), Maria Berenice R. Steffens

(UFPR, Curitiba), M. Geoffrey Yates (UFPR, Curitiba).

NATIONAL ADVISORY COMMITTEE

D. S. Andrade (IAPAR), J. I. Baldani (EMBRAPA Agrobiologia), R. Boddey (EMBRAPA

Agrobiologia), R. J. Campo (EMBRAPA Soja), A. A. Didonet (EMBRAPA Trigo), J.

Döbereiner (EMBRAPA Agrobiologia), A. A. Franco (EMBRAPA Agrobiologia), J. R. Jardim

Freire (UFRGS), F. M. S. Moreira (UFLa), L. A. de Oliveira (INPA), I. S. Schrank (UFRGS), J.

S. Souza (UFPR), S. M. Tsai (CENA), M. A. T. Vargas (EMBRAPA Cerrados).

INTERNATIONAL STEERING COMMITTEE

C. Elmerich (France)

H. Hennecke (Switzerland)

W. E. Newton (USA)

R. Palacios (Mexico)

F. O. Pedrosa (Brazil)

B. E. Smith (England)

I. Tikhonovich (Russia).

vii

INTERNATIONAL PROGRAM ADVISORY COMMITTEE

N. Amarger (France), M. Bazzicalupo (Italy), F. Bergersen (Australia), T. Bisseling (The

Netherlands), N. J. Brewin (UK), W. J. Broughton (Switzerland), B. Burgess (USA), F. D.

Dakora (South Africa), P. Dart (Australia), G. M. De Dretz (Uruguay), J. Dénarié (France), G.

Favelukes (Argentina), T. M. Finan (Canada), P. Gresshoff (USA), R. Haselkorn (USA), D.

Kahn (France), W. Klipp (Germany), A. Kondorosi (France), J. K. Ladha (Phillipines), D. B.

Layzell (Canada), A. B. Legocki (Poland), G. J. Leigh (UK), E. Martinez-Romero ( Mexico), M.

Megias (Spain), M. J. Merrick (UK), K. Minamisawa (Japan), Y. Okon (Israel), K. Pawlowski

(Germany), A. Pühler (Germany), B. Reinhold-Hürek (Germany), R. L. Richards (UK), L.

Seefeldt (USA), A. E. Shilov (Russia), H. P. Spaink (The Netherlands), G. Stacey (USA), J.

Thomas (India), R. N. F. Thorneley (UK), C. Vance (USA), D. Werner (Germany), S. S. Yang

(China), J. P. W.Young (UK).

SECRETARIAT

Mrs. J. Pie, Mrs. R. Prado, Mr. V. de Baura, Mr. A. Zimmermann, Mr. A. Copatti

(Departamento de Bioquímica da UFPR

MANAGEMENT

Mr. Laudálio Veiga Filho (General Manager - Provisuale Promoções

e Eventos e Programação Visual).

ACKNOWLEDGEMENTS

The editors would also like to express their gratitude to Vicki Newton for her expert advise and

guidance and to Liu Un Rigo, Emanuel Maltempi de Souza, Rose Adele Monteiro, Susan Hill,

Roseli Wassem and Maria Lucia Ishida for their help during the editing of this book.

DEDICATION

The editors dedicate this volume to

Dr. Johanna Döbereiner

and

Dr. João Rui Jardim Freire

in acknowledgement of their contributions

to nitrogen-fixation research in Brazil.

TABLE OF CONTENTS

PREFACE xxv

KEYNOTE LECTURENitrogen fixation in perspectiveW.E. NEWTON. 3

SECTION I: Biochemistry and ChemistryChemistry and biochemistry of nitrogenaseB.E. SMITH. 11

The mechanism of molybdenum nitrogenase: An overview.B.K. BURGESS. 13

Roles for nucleotides in nitrogenase catalysis.L.C. SEEFELDT, J.M. CHAN, W. WU, J.E. HUYETT. 19

Superoxide-dependent nitrogen fixation.C. HOFMANN-FINDEKLEE, D. GADKARI, O. MEYER. 23

Chemistry and biochemistry of nitrogenase (Part 1)R.N.F. THORNELEY, D.R. DEAN. 31

A 1.6 A resolution x-ray crystallographic study of Klebsiella pneumoniaeMoFe protein, Kp1S.M. MAYER, D.M. LAWSON, C.A. GORMAL, S.M. ROE, B.E. SMITH. 33

Roles of VnfX and NifX in FeV-co and FeMo-co synthesis in Azotobacter vinelandii.C. RÜTTIMANN-JOHNSON, P. RANGARAJ, C.R. STAPLES, V.K. SHAH ,P.W. LUDDEN. 35

Studies on the mechanism for the activation of iron and sulfur for formationof the nitrogenase metal centersD.R. DEAN, P. YUVANIYAMA, J.N. AGAR, M.K. JOHNSON. 37

Stopped-flow infra-red spectroscopy of carbon monoxide binding to functioningnitrogenaseR.N.F. THORNELEY, G.A. ASHBY, S.J. GEORGE . 39

Reductant-dependent ATP utilization during nitrogenase catalysis:Studies using Ti(III)A.C. NYBORG, J.A. ERICKSON, J.L. JOHNSON, G. D. WATT. 41

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Cofactor reactivity and models for cofactor reactionsG.J. LEIGH, R.L. RICHARDS. 43

Interactions of small molecules with isolated FeMocoC.J. PICKETT, S.P. BEST, T. LEGALL, C.A. GORMAL, S.K. IBRAHIM,B.E. SMITH, K. VINCENT. 45

Catalytic reactions with FeMoco in non-enzymatic surroundings (comparisonwith synthetic catalysts)A.E. SHILOV. 47

Catalytic behavior of isolated FeMo-cofactor of nitrogenase innon-protein surroundingsT.A. BAZHENOVA, M.A. BAZHENOVA, G.N. PETROVA, A.E. SHILOV. 49

Chemical nitrogen fixation: protonation of coordinated dinitrogen withcoordinated dihydrogen or bridging hydrosulfido ligands.M. HIDAI. 51

Mo, V and Fe complexes of tripodal sulfur-donor ligands as models fornitrogenase active sitesS.C. DAVIES, D.L. HUGHES, R.L. RICHARDS, J.R. SANDERS. 53

The use of chemical models to probe the mechanisms of substratereduction reactions of nitrogenasesC.A. HELLEREN, C.N. MCMAHON, G.J. LEIGH . 55

POSTER SUMMARIES. 57

SECTION II: Regulation of Nitrogen Fixation and Assimilation

Regulation of nif gene expression in free-living diazotrophs: recent advancesM.J. MERRICK, T. ARCONDEGUY, W.C. VAN HEESWIJK. 67

Activation of transcription by the sigma-54 RNA polymerase holoenzymeM.-T. GALLEGOS, M. CHANEY, P.CASAZ, W. CANNON, M.BUCK. 73

New mechanisms of bacterial gene regulation in a nitrogen-fixing phototroph.R.G. KRANZ, W.C. BOWMAN, N.R. SLOAN. 79

Regulation of nitrogen fixation and glutamine synthetase inHerbaspirillum seropedicaeE.M. SOUZA, M.G.YATES, R.WASSEM, M.BUCK, D. PERSUHN,E.M. BENELLI, G. KLASSEN, F.G.M. REGO, M.B.R. STEPHANS, L.U. RIGO,R.A. MONTEIRO, L.S. CHUBATSU, M. DRUMMOND, F.O. PEDROSA. 83

amtB is necessary for induced nitrogenase switch-off and ADP-ribosylationin Rhodobacter capsulatusA.F.YAKUNIN, P.C. HALLENBECK . 95

Regulation of Azotobacter vinelandii NifA activity by NifL: Role of PII-likeproteins in nitrogen sensingF. REYES-RAMIREZ, R. LITTLE, S. HILL, W. VAN HEESWIJK, R. DIXON . 97

Structural basis for signal transduction within the FixJ transcriptional activatorJ. SCHUMACHER, S. DA RE, J. FOURMENT, P. ROCHE, P. ROUSSEAU,B. TON-HOANG, D. KAHN. 99

Effect of redox status of dinitrogenase reductase on the regulation ofnitrogenase activity by reversible ADP-ribosylationC.M. HALBLEIB, Y.-P. ZHANG, B. ANTHARAVALLY, G.P. ROBERTS,P.W. LUDDEN . 101

Heterotrimerization of PII-like signalling proteins: Lessons from a comparativeanalysis between a cyanobacterial PII homologue and its proteobacterialcounterpartsK. FORCHHAMMER. 103

Role of PII protein as a signal of nitrogen level in Herbaspirillum seropedicaeE.M. BENELLI, E.M. SOUZA, A.C. BONATTO, M. BUCK, A. MOORE,M. HARPER, I. POLIKARPOV, F.O. PEDROSA. 105


87

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Analysis of bacterial gene expression during the late stages of the interaction betweenRhizobium etli CNPAF512 and Phaseolus vulgaris.J. MICHIELS, C. XI, B. DOMBRECHT, M. MORIS, G. DIRIX,J. VANDERLEYDEN

Functional analysis of the Bradyrhizobium japonicum RegSR two-componentregulatory proteinsR. EMMERICH, P. STREHLER, E. BAUER, H.-M. FISCHER, H. HENNECKE. 89

Role of Herbaspirillum seropedicae NifA domains on the expression ofnif genes.R.A. MONTEIRO, E.M. SOUZA, M.G. YATES, F.O. PEDROSA,L.S. CHUBATSU. 91

Characterization of an Azospirillum brasilense Tn5 mutant withenhanced nitrogen fixationC.A.G. BLAHA, L.F. REVERS, L.M.P. PASSAGLIA, J. FRAZZON,I.S. SCHRANK. 93

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Genetics of nitrogen fixation in Rhodobacter capsulatus: Ammonium andmolybdenum control of both nitrogenase systemsW. KLIPP, T. DREPPER, S. GROSS, B. MASEPOHL, K. RAABE, K.-U. RIEDEL,A.F. YAKUNIN, P.C. HALLENBECK. 141

Electron transport pathway to nitrogenase in Rhodobacter capsulatus:Rnf complex and its relatives in non-diazotrophsK. SAEKI, H. KUMAGAI. 143

Photosynthesis in Aeschynomene Bradyrhizobium sp.ORS278: Genetic analysisand role in symbiosisE. GIRAUD, L. HANNIBAL, C. CHAINTREUIL, J. LORQUIN, F. MOLOUBAS. MYLOVSKY, C. HURARD, C. BOIVIN, B. DREYFUS . 145

The concerted action of hydrogenases and nitrogenases in cyanobacteriaH. BOTHE, G BOISON, O. SCHMITZ. 147

Heterocyst pattern controlled by an inhibitory peptide in AnabaenaH.-S. YOON, J.W. GOLDEN. 149

The element nitrogen in the philatelic worldH. BOTHE 150

SECTION III: Taxonomy and Evolution of Diazotrophs

Taxonomy and evolution of diazotrophsJ.P.W. YOUNG. 153

Ecological, phylogenetic and taxonomic remarks on diazotrophs andrelated generaE. MARTÍNEZ ROMERO, J. CABALLERO-MELLADO, B. GÁNDARA,M.A. ROGEL, A. LÓPEZ-MERINO, E.T. WANG, L.E. FUENTES-RAMIREZI. TOLEDO, L. MARTINEZ, I. HERNANDEZ-LUCAS, J. MARTÍNEZ ROMERO. 155

Molecular evolution in diazotrophs: Do the genes agree?J.P.W. YOUNG. 161

Phylogeny of RhizobiaP. VAN BERKUM, J.J. FUHRMANN, B.D. EARDLY 165

Genetics of Nostoc in relation to host plant speciesU. RASMUSSEN, M.M. SVENNING. 171

Origin and evolution of nif genesR. FANI, S. CASADEI, P. LIO. 177

xv

Characterization of nitrogen–fixing bacteria containing molybdenum–independentnitrogenases from diverse natural environments.D. BETANCOURT, T.M. LOVELESS, P.E. BISHOP. 179

Biodiversity of rhizobia from a wide range of forest Leguminosae species in Brazil.F.M.S. MOREIRA.

181Biodiversity and selection of Sinorhizobium fredii strains for soybean inoculantsD.N. RODRÍGUEZ-NAVARRO, R. BELLOGIN, A.M. BUENDIA, M. CAMACHO,T. CUBO, L.M.O. CHUEIRE, A. DAZA, M.C. LYRA, R. ESPUNY, R. GOMEZ,M. HUNGRIA, H. MANYANI, M. MEGIAS, F.J. OLLERO, J.E. RUIZ-SAINZ,C. SANTAMARIA, C. SOUSA, F. TEMPRANO, S.S. YANG, J.C. ZHOU. 183

Taxonomy of the family AcetobacteraceaeL.E. FUENTES-RAMÍREZ, E. MARTÍNEZ-ROMERO, A. TAPIA-HERNÁNDEZ,R. BUSTILLOS, A. ROGEL, E.-T. WANG, T. JIMÉNEZ-SALGADO,J. CABALLERO-MELLADO . 185

Diversity in the bean nodulating rhizobial population of northwest ArgentinaO.M. AGUILAR, M.V. LOPEZ. 187


SECTION IV: Legume symbiosis

Control of nodulation in legume symbiosisG. STACEY. 211

Perception of lipo-chitooligosaccharidic Nod factors in the legume-rhizobia symbiosis:Studies on Medicago nod factor-binding sitesJ.-J. BONO, F. GRESSENT, A. NIEBEL, R. RANJEVA, J.V. CULLIMORE. 213

The use of GFP to study factors involved in the Lotus japonicus symbiosisH.P. SPAINK, H.R.M. SCHLAMAN, C. PACIOS-BRAS, A. ROUSSIS,J. STOUGAARD, N. STUURMAN. 219

Cell cycle control in root nodule organogenesisA. KONDOROSI, A. CEBOLLA, C. CHARON, F. FOUCHER, J. GYORGYEY,C. JOHANSSON, F. ROUDIER, C. SOUSA, J.M. VINARDELL, M. CRESPI,E. KONDOROSI. 223

Nod factor receptionG. STACEY, R.B. DAY, J. COHN, M. OKADA, Y. ITO, S. KOH, S. RAMU,T. UHM, D. KIM, D. COOK, N. SHIBUYA. 227

Functional characterization of a Krüppel-like zinc finger gene induced during noduledevelopmentF. FRUGIER, S. POIRIER, A. KONDOROSI, M..CRESPI. 231

xvi

Suppression of plant defence reactions in alfalfa cell cultures by Sinorhizobium melilotisurface carbohydratesK. NIEHAUS, U. ALBUS, R. BAIER, A. BECKER, K. SCHIENE, A. PÜHLER. 233

Invasion of alfalfa root nodules by the nitrogen-fixing bacterium Rhizobium melilotiG.R.O. CAMPBELL, B. PELLOCK, K. LEVIER, L.-X. WANG, S. M. BUSH,G.C. WALKER . 235

Structure and development of infection threadsN.J. BREWIN, E.A. RATHBUN, J.-P. WISNIEWSKI. 237

Unusual LPS structural features and symbiotic functionR.W. CARLSON, E. KANNENBERG, L.S. FORSBERG, B. JEYARETNAM. 239

Two Bradyrhizobium japonicum genes encoding putative sensor proteinsP. MÜLLER, E. MÜHLENCOERT. 241

POSTER SUMMARIES . 243

SECTION V: Structure and dynamics of diazotrophic and plant genomes

Genome structure of diazotrophsR. PALACIOS, M. MEGIAS. 269

Organizational, transcriptional and functional analyses of the Rhizobium sp.NGR234 genomeW.J. BROUGHTON, W.J. DEAKIN, M. FLORES, H.B. KRISHNAN, C. MARIE,P. MAVINGUI, R. PALACIOS, X. PERRET, V. VIPREY. 271

The symbiosis island of a Mesorhizohium strain that nodulates LotusC.W. RONSON, J. SULLIVAN, R. CRUICKSHANK, S. BROWN, R. ELLIOT,D. FLEETWOOD, A. HUBBER, N. MCCALLUM, K. MUIRHEAD, J. WEAVER,R. WEBBY, G. WIJKSTRA, J. TRZEBIATOWSKIZ, F. DE BRUIJN . 275

DNA reiteration in rhizobial genomes: How unusual is it?D. ROMERO, J. MARTÍNEZ-SALAZAR, E. ORTIZ, C. RODRÍGUEZ,E. VALENCIA-MORALES. 277

The symbiotic plasmid of Rhizobium etli: Sequence, analysis and comparisonG. DÁVILA, P. BUSTOS, M.A.CEVALLOS, J. COLLADO, V. GONZALEZ,J.C. HERNÁNDEZ, R. PALACIOS, V. QUINTERO. 279

Analysis of the symbiotic gene region of Bradyrhizobium japonicumM. GÖTTFERT, S. RÖTHLISBERGER, H. HENNECKE . 281

Multiple replicons in Azospirillum sp.C.C.G. MARTIN-DIDONET, L.S. CHUBATSU, E.M. SOUZA, M. KLEINA,F.G.M. REGO, L.U. RIGO, F.O. PEDROSA. . 283

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The MELILO EU project: Sequence of the 3.7 Mb chromosome of Sinorhizobium melilotiD. KAHN, F. BAYLOR-HUBLER, J. BATUT, A. BECKER, G. BOTHE, D. CAPLA,T. GODRIE, A. GOFFEAU, J. GOUZY, D. MASUY, T.M. POHL, D. PORTETELLE,A. PÜHLER, B. PURNELLE, P. THEBAULT, M. VANDENBOL, S. WEIDNER,F. GALIBERT. 285


SECTION VI: Plant genes involved in symbiosis

Towards the understanding of the structure and function of sym genesI.A. TIKHONOVICH. 303

Functional genomics and genetic analysis of nodulation of soybean andLotus japonicusP.M. GRESSHOFF, A.E. MEN, A. HUSSAIN, Q. JIANG, D. LOHAR,I.L. ANDERSSEN, C. ZAHARI, C.–A. VASILE, M. SPENSER, L. STILLER. 305

A region on the upper arm of chromosome 5 of Medicago truncatula is highlysyntenic to the SYM2 region of peaR. GEURTS, G. GUALTIERI, O. KULIKOVA, D.-J. KIM, D. R. COOK,T. BISSELING. 309

Plant genes controlling development of two pea (Pisum sativum L.)endosymbiotic systemsI.A. TIKHONOVICH, B. BOESTEN, A.Y. BORISOV, L.M. JACOBI,A.S. KUKALEV, V.K. LEBSKY, E.V. MORZHINA, J. OLIVARES,N.E. PETROVA, U.B. PRIEFER, V.I. ROMANOV, J. SANJUAN,J.M. SANJUAN-PINILLA, A.F. TOPUNOV, V.E. TSYGANOV,V.A. VOROSHILOVA. 311

Modulation of ammonium assimilation in transgenic legumes during the symbiosis withRhizobiumG. HERNÁNDEZ, M. LARA, E. CÓRDOBA, E. DIEGO-GARCÍA, S. CHICHKOVA. 313

An effort towards the map-based cloning of a gene conditioning non-nodulationtrait in tetraploid Medicago sativaG.B. KISS, P. KALÓ, A. KERESZT, S. MIHACEA, G. ENDRE. 315

Genetic linkage mapping and location of genomic regions associated with nodulationand bacterial diseases in Phaseolus vulgaris LS. M. TSAI, L.E.A. CAMARGO, R.L. BOSCARIOL, A.A. SOUZA, D.H. MOON,A.V.O.FIGUEIRA, P.GEPTS. 317

Two subclasses of yellow lupine PR10 proteins and their possible functionduring the symbiosis developmentM.M. SIKORSKI, L. HANDSCHUH, J. BIESIADKA, A B. LEGOCKI. 319

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Early nodulin genes as markers of various developmental stages during nodulation ofMedicago truncatula by Sinohirizobium melilotiR. MATHIS, F. DE BILLY, M.C. BOUDET, V. LEGER, P.GAMAS. 323

Early symbiotic functions of Sesbania rostrataM. HOLSTERS, S. LIEVENS, W. VAN DE VELDE, M. CATURLA-GOÑI,W. D’HAEZE, M. GAO, S. GOORMACHTIG. 325

Characterization and analysis of symbiotic genes in alfalfa and white sweetcloverA.M. HIRSCH, N.A. FUJISHIGE, R.E. SATTERTHWAITE, A. HERNANDEZ,W.M. KARLOWSKI. 327

Nin, a developmental regulator of root nodule initiation in Lotus japonicusL. SCHAUSER, A. ROUSSIS, A. NIELSEN, J. STOUGAARD. 329

Exploring the genetic potential of rice for forming symbiotic associations with rhizobiaP.M. REDDY, R.J. HERNANDEZ-OANE, H. KOUCHI, G. STACEY, J.K. LADHA. 331


SECTION VII: Metabolism in symbiosomes

Symbiosome metabolism in legume nodulesD.A. DAY. 349

Assimilation of fixed nitrogen studied by membrane vesicle transport andin vivo 15N NMR spectroscopy in peaL. ROSENDAHL, A. RUDBECK, A.M. SCHARFF, P. MOURITZEN. 351

Alanine excretion from Bradyrhizobium japonicum bacteroidsD.W. EMERICH, J.K. WATERS, B.L. HUGHES, L.C. PURCELL,K.O. GERHARDT, T.P. MAWHINNEY. 355

Carbon metabolism and symbiotic needs of root nodule bacteriaM. MITSCH, A. COWIE, R. VOEGELE, T.M. FINAN. 359

Molecular biology of the peribacteroid membraneM.K. UDVARDI , G. DE BRUXELLES, D. DAY, S. FRUEND, F. GREIL, B. KAISER,D. LAVER, S. PANTER, U. SIMON, R. THOMSON, B. TREVASKIS. 365

Oxygen regulation and adenylates in legume nodulesD.B. LAYZELL. 367

The putative bd-type terminal oxidases of Bradyrhizobium japonicumE. ARSLAN, A. KANNT, R. ZUFFEREY, F. BAUMANN, L. THONY-MEYER,H. HENNECKE. 369

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Ammonium and alanine are the primary nitrogen secretion products of pea bacteroidsP.S. POOLE, D. ALLAWAY, E. LODWIG, T. WHEELER. 371

Regulation of assimilation during symbiosome differentiation in Rhizobium etliE.J. PATRIARCA. 373

The role of rhizobitoxine in nodulation by Bradyrhizobium elkaniiK. MINAMISAWA, T. YASUTA, K. YUHASHI, N. ICHIKAWA, H. EZURA,N. NUKUI. 375

Polyhydroxybutyrate in nitrogen-fixing symbiosesL. COPELAND, L. FENG, C. TABRETT 377


SECTION VIII: Associative and endophytic associatoins

Recent progress in characterization of associative and endophytic diazotrophsand their influence on host plant growthC. KENNEDY. 395

Biological nitrogen fixation (BNF) in non-leguminous plants: The role of endophyticdiazotrophsJ.I. BALDANI, A.L.M. OLIVEIRA, S.L. GUIMARAES, V.L.D. BLADANI,F.B. REIS, JR., L.G. SILVA, V.M. REIS, K.R.S. TEIXEIRA, J.DÖBEREINER. 397

Analysis of genes for nitrogen fixation and studies of plant growth enhancementIn the diazotrophic endophyte of sugarcane, Acetobacter diazotrophicusC. KENNEDY, S. LEE, M. SEVILLA, D. MELETZUS, N. GUNAPALA,A. GARDIOL, S. DAVIDSON. 401

Interactions of diazotrophic Azoarcus spp. with riceB. REINHOLD-HUREK, J. DÖRR, T. EGENER, D. MARTIN, T. HUREK. 405

Associative and endophytic symbiosisM. BAZZICALUPO, Y. OKON. 409

Molecular approaches for the investigation of the diversity and localization of diazotrophicbacteriaA. HARTMANN, B. ECKERT, M. STOFFELS, F.L. OLIVARES, O. WEBER,V. REIS, V.L.D. BALDANI, J.I. BALDANI, M. SCHLOTER, G. KIRCHHOF. 411

Endophytic establishment of diazotrophic bacteria in sugar cane plantsF.L. OLIVARES, E.K. JAMES. 413

Involvement of extracellular components in the aggregation of Azospirilium brasilenseS. BURDMAN, E. JURKEVITCH, Y. OKON. 415

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The cytochrome (cytochrome c) terminal oxidase of Azospirillum brasilense SP7K. MARCHAL, J. SUN, V. KEIJERS, H. HAAKER, J. VANDERLEYDEN. 417

Diversity of endophytic diazotrophs and mechanism of endophytic colonizationin deep water riceS.C. VERMA, J.K. LADHA, A.K. TRIPATHI. 419


SECTION IX: Actinorhizal associations

Frankia and actinorhizal plantsK. PAWLOWSKI. 451

Sugar partitioning in nitrogen-fixing root nodulesK. PAWLOWSKI, P. WABNITZ, M. RAMENSKAIA. 453

Flavan-containing cells delimit Frankia infected compartments inCasuarina glauca nodulesL. LAPLAZE, H.GHERBI, T. FRUTZ, K. PAWLOWSKI, C. FRANCHE,J.J. MACHEIX, F. AUGUY, D. BOGUSZ, E. DUHOUX . 455

Casuarina-Frankia symbioses – molecular studies of FrankiaA. SELLSTEDT, U. MATTSSON, F. TAVARES, C. LUNDBERG. 457

Distribution and phylogeny of Gymnostoma spp. infective Frankia strains inNew CaledoniaE. NAVARRO, T. JAFFRE, P. NORMAND 459

Specificity in Discaria - Frankia symbiosesL.G. WALL, E. CHAIA, C. VALVERDE, G. LUCKI. 461

Novel nodule-specific glycine- and histidine-rich proteins expressed in the zone ofinfection of actinorhizal nodules may be multimeric metal-binding proteinsS.V. DOBRITSA, C.M. MAILLET, B.C. MULLIN. 463

POSTER SUMMARIES 465

SECTION X: Environmental stress

Physiological roles of betaines in Sinorhizobium meliloti and other members of theRhizobiaceae subjected to salt stressD. LE RUDULIER, E. BONCOMPAGNI, A. LAMBERT, M. OSTERAS, L. DUPONTK. MANDON, J.C. TRINCHANT, M.C. POGGI. 473

Genetic circuits involved in the response of root nodule bacteria to low pHR.P. TIWARI, W.G. REEVE, M.J. DILWORTH, A.R. GLENN. 475

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Persistence and competitiveness of Or191-like rhizobia and Sinorhizobium meliloti(Sme) strains in an acidic soil of ArgentinaL.J. BALAGUÉ, M.F. DEL PAPA, M. PISTORIO, A. PERTICARI, A. LAGARES. 477

A requirement for the iron regulated outer membrane protein FegA in theBradyrhizobium japonicum/ soybean symbiosisE. BONCOMPAGNI, M.L. GUERINOT. 479

2D SDS PAGE of proteins involved in acid tolerance in Rhizobium tropici CIAT 899H.L. STEELE, P. VINUESA, M. STRÜMPELER, R. SCHMID, D. WERNER. 481

A mutation in the guaB gene of R. tropici affects thermal tolerance and bean nodulationP.M. RICCILLO, M.M. COLLAVINO, R. ENGLAND, F.J. DEBRUIJN,O.M. AGUILAR. 483


SECTION XI: Biological nitrogen fixation in sustainable agriculture

Biological nitrogen fixation: Phosphorus - a critical future need?C.P.VANCE, P.H. GRAHAM, D.L. ALLAN. 509

The Brazilian experience with the soybean (Glycine max) and common bean (Phaseolusvulgaris) symbiosesM. HUNGRIA, M.A.T. VARGAS, R.J. CAMPO, L.M.O. CHUEIRE,D. DE S. ANDRADE . 515

Quantification of biological nitrogen fixation in agricultural systemsM.B. PEOPLES, D.F. HERRIDGE. 519

Putting legume to work in cropping systems of southern AfricaK.E. GILLER, S. MPEPEREKI, P. MAPFUMO, P. CASASA, W. SAKALA,H. PHOMBEYA, O. ITIMU, G. CADISCH, R.A. GILBERT, S.R. WADDINGTON. 525

Measuring nitrogen fixation by treesR.M. BODDEY, P.J. DART, M.B. PEOPLES. 531

Benefit of legume-fixed N in crop rotations under zero tillageB.J.R. ALVES, L. ZOTARELLI, W.A.R. LARA-CABEZAS, E. TORRES,M. HUNGRIA, S. URQUIAGA, R.M. BODDEY 533

Self regulation of biological fixation of tree legumes in a forest successionof the eastern AmazonL. PAPARCÍKOVÁ, A. THIELEN-KLINGE, P.L.G. VLEK. 535

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Multipurpose legumes of the tropical genus Crotalaria are associated withunusual rhizobiaA. SY, P. JOURAND, E. GIRAUD, I. NDOYE, P. DE LAJUDIE, C. BOIVIN-MASSON,R. SAMBA, M. NEYRA, A. WILLEMS, M. GILLIS, B. DREYFUS. 537

Nitrogen fixation by forage legumes as a driving force behind the recuperationand improvement of soil quality in tropical agricultural systems:Opportunities for wider use of forest legumes?R.J. THOMAS. 539


SECTION XII: Soil reclamation and remediation

The importance of biological nitrogen fixation on land rehabilitationA. A. FRANCO, E.F.C. CAMPELLO, S.M. DE FARIA, L.E. DIAS. 569

Soil biotests and phyto remediation with the legume-Rhizobium symbiosesD. WERNER, H. NEUMANN, H. STEELE, A. WETZEL, R. MAGASHEVA,R. PLISAK. 571

Can aquatic plants remove heavy metals from industrial wastes and sludges effectively?E. TEL-OR, G. NAHUM, N. SHOEL, R. BENAROYA, N. LAVID, E. SOKOLOVKSI,Z. KEILIN, T. SCHOR, Y. LIBAL. 575

Bioremediation of metal-contaminated soilsM.J. SADOWSKY. 577

Host-plant factors in the adaptation of indigenous African legumes tolow pH soilsF.D. DAKORA, A. SPRIGGS, R.C. NYEMBA, S.B.M. CHIMPHANGO. 579

A new remediation system for heavy metals using leguminous plantand rhizobia symbiosisY. MUROOKA, M. GOYA, S.-H. HONG, M. HAYASHI, H.ONO,M. TACHIMOTO, N. HIRAYAMA. 581


SECTION XIII: Strain selection and inoculation technology

Legume inoculation in cerrado soilsM.A.T. VARGAS, M. HUNGRIA. 587

Impact of new technologies on inoculation by diazotrophsG. MARTINEZ-DRETS, S. BATISTA, S. CASTRO. 589

Antibiosis as a means to enhance nodulation competitiveness:Beyond the model systemE.A. ROBLETO, A.J. SCUPHAM, E.W.TRIPLETT.

Horizontal gene transfer in rhizobia: Ecological implicationsJ. SANJUAN, J.A. HERRERA-CERVERA, J.M. SANJAUAN-PINILLA,S. MUNOZ, J. NOGALES, J. OLIVARES.

Plant-biotin a possible signature compound for Sinorhizobium melilotiduring alfalfa root colonizationW.R. STREIT.

Importance of molybdenum and cobalt to the biological nitrogen fixationR.J. CAMPO, U.B. ALBINO, M. HUNGRIA.

POSTER SUMMARIES.

SECTION XIV: Nitrogen fixation imperatives for the century

Prospects for engineering nitrogen-fixing photosynthetic eukaryotes.R. DIXON, Q. CHENG, M. DOWSON-DAY, A. DAY.

Nitrogen fixation in rice: Objectives and achievementsP.M. REDDY, J.K. LADHA.

Genetic transformation of legumesC.A. ATKINS, P.M.C. SMITH.

Can the study of endomycorrhizae open new avenues of reserarch in symbioticnitrogen fixation?C. GOUGH, P. BONFANTE, J. DÉNARIÉ.

AUTHOR INDEX

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591

593

595

597

599

635

641

653

659

647

PREFACE

The 12th International Congress on Nitrogen Fixation was held in Foz do Iguaçu, Paraná State,Brazil, from the 12th to 17th of September, 1999. This volume constitutes the proceedings ofthis Congress and represents a compilation of the presentations by scientists from 38 countrieswho came to discuss the progress made, to exchange views and to collaborate. Since the firstmeeting in Pullman, Washington, USA, in 1974, this series of Congresses has been held threetimes in the United States, four times in Western Europe, in Australia, Mexico and Russia, andnow for the first time in South America. Brazil was a most appropriate choice because Brazilianagriculture is especially dependent on biological nitrogen fixation. An important example is thesoybean crop. The 30 million metric tons of grain produced each year relies solely on symbioticnitrogen fixation. N-fertilizers are expensive in Brazil and, in the absence of the biologicalprocess, this crop would require 4-5 M tonnes of N at an annual cost of almost 1 billion dollars.Crop rotation, soil reclamation, forestry, forage mixed cropping, intercropping with grasses andlegumes, associative nitrogen fixation with non-legume crops, such as sugarcane, and otheragricultural practices in Brazil are also dependent on biological nitrogen fixation. The Brazilianexperience with biological nitrogen fixation is a paradigm to be followed by all countries wherenitrogen fertiliser is expensive or where present day environmental concerns demand a feasible,efficient and inexpensive alternative to chemical N-fertilizers.

In the biochemistry section, the analysis of the structure of the MoFe protein of Klebsiellapneumoniae at a resolution of 1.6 angstroms was described as was the complex between thealtered Azotobacter vinelandii Fe protein which remains locked in the MgATPconformation in the absence of MgATP, and the MoFe protein. The intriguing possibility wasraised that the Fe protein can be reduced by two electrons and transfer them both during a singleencounter with the MoFe protein thereby improving energy expenditure with only two MgATPshydrolysed per two electrons transferred. Other advances include the study of the FeMoco innon-protein surroundings, involving the binding of inhibitors and the reduction of substrates.These studies are paralleled by FTIR analysis of CO binding to the MoFe protein. Theregulation of nif gene expression and nitrogen metabolism attracted a large number ofcontributions. Major advances in regulation have centered on the activation of the nifA gene andthe activity of the NifA protein. Regulation of activity by NifL involves the GlnK protein ineither a positive or negative manner, depending on the organism. The N-terminus of theHerbaspirillum protein inhibits the binding of the central plus C-terminal domain to the nifHpromotor, whereas nifL probably prevents NifA interacting with the RNA polymerase. PII ofHerbaspirillum, which activates NifA, has a structure similar to the GlnK of Escherichia coli.Autoactivation of the nifA gene in Herbaspirillum is inhibited by IHF, which acts positively andnegatively, to regulate nifA expression. The role of the protein has been studied in depth.

A big shift has been made from genes to genomes. The sequence of the sym plasmid of NGR234 has been completed as has the genome of Rhodobacter capsulatus. Other genomes underinvestigation are those of Sinorhizobium meliloti, Bradyrhizobium japonicum, the symbiosisisland of a Mesorhizobium loti and the symbiotic plasmid of Rhizobium etli. Plant genomes thathave been analyzed include Medicago truncatula, Lotus japonicus and soybean.

Interest persists in the associative and endophytic bacteria and evidence is growing that theendophytic bacteria, such as Acetobacter diazotrophicus, Herbaspirillum and Azoarcus species,

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colonize intercellular spaces in the cortex and also xylem vessels. The range of plant specieshosting Azospirillum, Herbaspirillum and Burkholderia, as well as undefined species hasincreased. Clear evidence has been obtained that nitrogenase is expressed inside sugarcaneplants harbouring Acetobacter diazotrophicus and inside rice harbouring Azoarcus spp. Becausenitrogenase is expressed under these conditions, biological nitrogen fixation presumablycontributes to bacterial growth and possibly to plant growth. However, the positive contributionof these organisms to plant growth goes beyond just nitrogen fixation.

Nod factors (LCO’s) were the major focus concerning the control of nodulation in the legumesymbiosis. Purified Nod factors, together with expression of the enod40 gene, activate the innercortical cells for the cell division necessary for nodule formation. Differentiation ofmeristematic cells involves arrest of cell division, an increase in cell size and infection bybacteroids. A plant cell regulator gene ccs52 controls cell-division arrest and is essential fornodule formation. Lectin-like NTPases appear to be necessary for nodulation, possibly as Nodfactor-binding proteins. A key issue in the session on metabolism of symbiosomes concernedalanine, rather than ammonium, as the major N exporter from the nodule to the plant. It is likelythat either ammonium or alanine can be excreted, depending on the physiological state of thebacteroid. Interest continues to be high regarding both the nodule malic enzyme and theperibacteroid membrane.

New molecular tools are enabling progress in the study of molecular phylogeny and biologicaldiversity. Several studies show similarities between species and other species andgenera not reported as fixers. Much effort is being expended in trying to unravel andcomprehend the biodiversity of diazotrophic bacteria.

Agricultural practices emphasized the benefits of nitrogen fixation for improving soil fertilityand increase yield, for playing an important role in the sustainable management of agriculturalsystems, and in soil reclamation. Phosphorus remains a key nutrient to improve theeffectiveness of the nitrogen-fixation process and, therefore, the important role played bymycorrhizae was emphasized. Approaches using the inoculation of selected strains,improvement of inoculation technology, co-inoculation of microorganims and several otherpractices are being successfully used in several countries.

A final session was devoted to predictions of future research directions in nitrogen fixation forthe 21st century. These included the possibility of establishing active nitrogen fixation in plants,the establishment of nitrogen-fixing symbioses with rice, and the use of transgenic legumes tocombat environmental stress, including herbicide tolerance and resistance to pests and diseases.

We want to express our deep thanks to all those who worked hard and long to help us in theorganization of this Congress. We want to emphasize our gratitude for the financial support thatwe received from all the institutions that believe in our work. Finally, we want to thank ourfamilies and friends for their moral support during the last two years.

Curitiba, September 1999

Fábio O. PedrosaMariangela Hungria

M. Geoffrey YatesWilliam E. Newton

NITROGEN FIXATION: FROM MOLECULES TO CROP …978-0-306-47615-0/1.pdf · Nitrogen Fixation: From Molecules to Crop Productivity Proceedings of the International Congress on Nitrogen

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