BIOLOGICAL NITROGEN FIXATION, SUSTAINABLE AGRICULTUREAND THE ENVIRONMENT

Current Plant Science and Biotechnology in AgricultureVOLUME 41

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

Scientific Advisory BoardJ. Hamblin, Research Director, Export Grains Centre Ltd., 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 previousvolumes in the series dealt with plant science and biotechnology, the aim is now to also includevolumes dealing with animals science, food science and microbiology. While the subject matter willrelate more particularly to agricultural applications, timely topics in basic science and biotechnologywill also be explored. Some volumes will report progress in rapidly advancing disciplines throughproceedings of symposia and workshops while others will detail fundamental information of anenduring nature that will be referenced repeatedly.

The titles published in this series are listed at the end of this volume.

Biological Nitrogen Fixation,Sustainable Agriculture andthe EnvironmentProceedings of the 14th International Nitrogen Fixation Congress

Edited by

YI-PING WANGCollege of Life Sciences,Peking University,Beijing, China

MIN LINInstitute of Biotechnology,Chinese Academy of Agricultural Sciences,Beijing, China

ZHE-XIAN TIANCollege of Life Sciences,Peking University,Beijing, China

CLAUDINE ELMERICHInstitut des Sciences du Végétal,

and

WILLIAM E. NEWTONVirginia Polytechnic Institute and State University,Blacksburg, VA, USA

CNRS-UPR2355, Gif-sur-Yvette and Institute Pasteur, Paris, France

Library of Congress Cataloging-in-Publication Data

Printed on acid-free paper

ISBN-10 1-4020-3569-1 (HB) ISBN-10 1-4020-3570-5 (e-book) ISBN-13 978-1-4020-3569-1 (HB) ISBN-13 978-1-4020-3570-5 (e-book)

Published by Springer,P.O. Box 17, 3300 AA Dordrecht, The Netherlands.

All Rights Reserved© 2005 SpringerNo part of this work may be reproduced, stored in a retrieval system, or transmittedin any form or by any means, electronic, mechanical, photocopying, microfilming, recordingor otherwise, without written permission from the Publisher, with the exceptionof any material supplied specifically for the purpose of being enteredand executed on a computer system, for exclusive use by the purchaser of the work.

Printed in the Netherlands.

www.springeronline.com

14TH INTERNATIONAL CONGRESS

ON NITROGEN FIXATION

was organized by

NATIONAL LABORATORY OF PROTEIN ENGINEERING AND PLANT GENETIC ENGINEERING,

COLLEGE OF LIFE SCIENCES, PEKING UNIVERSITY BIOTECHNOLOGY RESEARCH INSTITUTE, CHINESE ACADEMY OF

AGRICULTURAL SCIENCES

and supported by

Peking University Ministry of Education, P. R. China

National High Technology Research and Development Program of China (863 Program)

The National Key Fundamental Research Program of China (973 Program) National Natural Science Foundation of China

UNESCONational Laboratory of Protein Engineering and Plant Genetic Engineering

Chinese Society of Biotechnology Chinese Society for Microbiology

Sichuan Habio Bioengineering Co., Ltd Shenfeng Biotech Engineering Co., Ltd of Angang Industry

14TH INTERNATIONAL CONGRESS

ON NITROGEN FIXATION

was organized by the following committees

National Organizers

National Laboratory of Protein Engineering and Plant Genetic EngineeringCollege of Life Sciences, Peking University

Biotechnology Research Institute, Chinese Academy of Agricultural Sciences

National Organizing Committee

Y.P. Wang, College of Life Sciences, Peking University, Beijing, China M. Lin, Institute of Biotechnology, CAAS, Beijing, China

Y.X. Jing, Institute of Botany, CAS, Beijing, China

National Program Committee

S.J. Shen (S.C. Shen), Shanghai J.L. Tang, Nanning J.L. Li, Beijing Z.X. Tian, Beijing Q.R. Cai (K.R. Tsai), Xiamen Y.P. Wang, Beijing S.F. Chen, Beijing B. Wu, Nanning W.X. Chen, Beijing X.T. Wu, Fuzhou C. Ge, Beijing L. Xu, Shanghai J.F. Huang, Beijing J. Q. Xu, Changchun Y.X. Jing, Beijing S.S. Yang, Beijing F.D. Li, Wuhan G.Q. Yu, Shanghai J. Li, Beijing Y.X. Yu, Chongqing J.D. Li, Beijing C.G. Zhang, Shenyang Z.T. Li, Beijing X.P. Zhang, Chendu M. Lin, Beijing Z.Z. Zhang, Shenyang Q.S. Ma, Nanning J.D. Zhao, Beijing Y.S. Qiu, Guangzhou J.C. Zhou, Wuhan S.H. Shen, Beijing J.B. Zhu, Shanghai Y.X. Zhu, Beijing

14TH INTERNATIONAL CONGRESS

ON NITROGEN FIXATION

International Steering Committee

W.E. Newton, USA F.O. Pedrosa, BrazilR. Palacios, Mexico A. Pühler, GermanyC. Elmerich, France I. Tikhonovich, RussiaH. Hennecke, Switzerland T. Finan, CanadaB.E. Smith, UK Y.P. Wang, China

International Program Advisory Committee

N. Boonkerd, Thailand K.A. Malik, PakistanW. Broughton, Switzerland E. Martinez-Romero, MexicoM. Buck, UK K. Minamisawa, JapanE.C. Cocking, UK M. O'Brian, USAF. Dakora, South Africa M. Rahman, BangladeshH. Das, India B. Rolfe, AustraliaF. de Bruijn, France C. Ronson, New ZealandR. Dixon, UK T. Ruiz-Argueso, SpainZ.M. Dong, Canada S.J. Shen (S.C. Shen), ChinaD. Emerich, USA H.P. Spaink, The NetherlandsR. Fani, Italy G. Stacey, USAP. Gresshoff, Australia S. Tabata, JapanR. Haselkorn, USA Nguyen Thanh Hein, VietnamR. Holm, USA R. Thorneley, UKC. Kennedy, USA C. Vance, USAA. Kolb, France D. Werner, GermanyJ.K. Ladha, Philippines F. O'Gara, Ireland P. Ludden, USA K. Pawlowski, Germany

Greetings from Prof. San Chiun Shen

“Progress in Nitrogen Fixation Research relies on the integration of knowledge from different fields, plus scientific collaboration and a free

exchange of views between scientists. Accumulation of research data over many years is the guarantee of innovation. To be at the top in science does

not mean to be at the front of the stage. The best work will always be recognized, and will lead us all along the path of truth and understanding.

This is the driving force of science.”

National Laboratory of Plant Molecular Genetics, Institute of Plant Physiology and Ecology, Shanghai Institutes of Biological Sciences, Chinese Academy of Sciences,

Shanghai, P. R. China

xi

TABLE OF CONTENTS

PREFACE…………………………………………………………………………..xxiii

DEDICATION…………………………………………………………………………..1

In Memory of Professor Jia-Xi Lu Xintao Wu…………………………………………………………………….……....... 3

A Life Affair with Rhizobia – In Memory of Professor Chen Hua-Kui, Academician of the Chinese Academy of Sciences Fu-Di Li………………………………………………………….…………….….….…7

KEYNOTE ADDRESS………………….………………………………………….….11

Sustainable Agriculture/Forestry and Biological Nitrogen Fixation Dietrich Werner, Suresh K. Mahna, Vertica Mahobia, B.N. Prasad, Jose

M. Barea, Heidemarie Thierfelder, Peter Müller and Pablo Vinuesa………………….13

SECTION I. NITROGENASE: THE ORIGIN, THE ENZYMES,

AND THE CHEMISTRY………………..……………………………………………19

On the Origin and Evolution of nif Genes Renato Fani……………………………………………………………………………..21

Recent Progress in the Biochemistry of Mo-Nitrogenase William E. Newton……………………………………………………………………...25

MgATP Binding and Hydrolysis in Nitrogenase Catalysis Sanchayita Sen and John W. Peters…………...………………………………………..29

Structural Modeling and Chemical Simulation of the FeMoco Active Center of Nitrogenase Shaowu Du, Qiutian Liu, Beisheng Kang, Changren Chen and Rong Cao……………35

A Non-Metal System for Nitrogen Fixation Utilizing Buckminsterfullerene Yoshiaki Nishibayashi, Sakae Uemura, Shin-ichi Takekuma and Zen-ichi Yoshida…....39

Chemical Simulation of the Molybdenum Coordination Mode in the FeMo Cofactor by a Series of Molybdenum- or Tungsten-Polycarboxylate Complexes with Sulfide Bridges Dong-Mei Li, Ji-Qing Xu, Yong-Heng Xing, Ying-Jie Lin and Xiao-Hua Zhou………..41

xii

Activation of Homocitrate Ligand of FeMo-Cofactor as Mimicked by the Coordination of Homocitrato and its Homologues Z.H. Zhou, S. Ya Hou, Z. Xing Cao, Y. Fu Deng, H. Lin Wan, and K. Rui Tsai….…..…43

Poster Summaries………………………………………………………………………46

SECTION II. GENETICS AND REGULATION IN HETEROTROPHS

AND PHOTOSYNTHETIC BACTERIA……………………………………………51

Genetic Regulation of Nitrogen Fixation: Integration of Multiple Signals Richard Little, Isabel Martinez-Argudo, Neil Shearer, Philip Johnson and

Ray Dixon………………………………………………………………………………53

The Structural Basis for nif Gene Activation S. R. Wigneshweraraj, P. C. Burrows, P. Bordes, J. Schumacher, M. Rappas,

R.D. Finn, W.V. Cannon, X. Zhang and M. Buck……………………………………….59

Heterocyst Differentiation and Nitrogen Fixation in Anabaena

Robert Haselkorn……………………………………………………………………….65

Signal Transduction in Heterocyst Differentiation of the CyanobacteriumAnabaena sp. PCC 7120 Yinhong Zhao, Yunming Shi, Guohua Yang, Ying Zhang, Xu Huang and

Jindong Zhao…………………………………………………………………………...69

P Signal Transduction in Cyanobacteria: Novel Mechanisms of Global Nitrogen Control Karl Forchhammer, Annette Heinrich, Nicole Kloft, Mani Maheswaran and

Ulrike Ruppert………………………………………………………………………….73

Molecular Signal of Nitrogen Starvation, Insight from the Cyanobacterium Anabaena PCC 7120 Cheng-Cai Zhang, Sophie Laurent and Sylvie Bédu……………………………….…..77

Electron Transport to Nitrogenase in Rhodospirillum rubrum

Tomas Edgren and Stephan Nordlund………………………………………………….79

Regulation of Nitrogen Fixation in the Photosynthetic Bacterium Rhodospirillum rubrum: Roles of GlnD and P ProteinsYaoping Zhang, Edward L. Pohlmann and Gray P. Roberts………………………..…83

xiii

Functional Analysis of the NifA GAF Domain of Azospirillum brasilense:

Effect of Tyr-Phe Mutations in NifA and Interaction of the Mutated Proteins with GlnB using the Yeast Two-Hybrid System Sanfeng Chen, Li Liu, Xiaoyu Zhou, Claudine Elmerich and JiLun Li……………..…87

Effects of 2-Oxoglutarate and the P Homologues nifI1 and nifI2 on Nitrogenase Activity in Cell Extracts of Methanococcus maripaludis

J.A. Dodsworth and J.A. Leigh ………………………………………………………...89

Mechanism of Regulation of Nitrogen Fixation by NifLA in Pseudomonas

Stutzeri A1501 Zhihong Xie, Jie Pan, Shuzheng Ping, Ming Chen, Guoying Wang, Yi Yang,

Min Lin and Claudine Elmerich………………………………………………………..91

Rhizobial FixL/FixJ System is a Paradigmn of the Two-Component Signal Transducing Systems: A Two-Cylinder Reciprocating Engine Model Of Phosphorylation Reactions Hiro Nakamura………………………………………………………………………....93

Poster Summaries…………………………………………………………………........95

SECTION III. FROM GENOMES TO FUNCTION…………………………....109

A Report on the Genome of Herbaspirillum seropedicae Strain Z78 Fabio O. Pedrosa and Genopar Consortium………………………………………….111

Functional Analysis of Genes of Unknown Functions in Sinorhizobium

meliloti 1021J. Cheng, J. Fowler, A. Cowie, R. Zaheer, C. Patten, C. Sibley, P. Chain,

Z. Yuan, C. Baron, T. Charles, B. McCarry, P. Summers, J. Xu, E. Weretilnyk,

G.B. Golding and T.M. Finan………………………………………………………….115

What Can Bacterial Genome Research Teach Us about Iron Uptake in Sinorhizobium meliloti?A. Pühler, A. Becker, J. Buhrmester, T.-C. Chao, S. Rüberg and S. Weidner………….119

Structural and Functional Genome Analysis of Lotus japonicus and Mesorhizobium loti

S. Sato, T. Kaneko, Y. Nakamura, E. Asamizu, T. Kato and S. Tabata…………...……123

xiv

Analyzing a Sinorhizobium meliloti 1021 ORFeome in a Functional Genomics Platform Brenda K. Schroeder, Brent L. House, Michael W. Mortimer, Scott C. Maloney,

Casey A. Taylor, Kristel L. Ward, Hope T. Ziemkiewicz, Scott Clark, John

J. Bovitz, Hao Jin, Svetlana Yurgel and Michael L. Kahn…………………………….127

Functional Genomic Analysis of the SDR Family in Sinorhizobium meliloti

Sirin Adham, Asha Jacob, David Capstick, Trevor Charles…………………………..129

Proteome Analysis on Bacteroid Differentiation of Bradyrhizobium

Japonicum USDA 110 Shigeyuki Tajima, Le T-P Hoa, Rie Hamaguchi and Mika Nomura…………………..131

Global Changes in Gene Expression of S. meliloti 1021 Nodule Bacteria in nifA and nifH Mutant Background Zhe-Xian Tian, Hua-Song Zou, Jian Li, Yuan-Tao Zhang, Guan-Qiao Yu,

Silvia Rüberg, Anke Becker and Yi-Ping Wang……………………………………….133

Toward Deciphering the Genome of Frankia alni Strain ACN14a. P. Normand., S. Felix, N. Alloiso, J. Marechal, C. Lavire, A.M. Berry,

B.C. Mullin, J., Tomkins, N. Choisne, N. Demange, Y.C. Truong Cong,

A. Coulloux, D. Vallenet, S. Cruveiller, C. Médigue…………………………………..137

Poster Summaries……………………………………………………………………..139

SECTION IV. ESTABLISHMENT OF SYMBIOSIS AND NODULE

FUNCTION…………………………………………………………………………..145

Cell Cycle and Symbiosis Adam Kondorosi, José Maria Vinardell, Toshiki Uchiumi, Peter Mergaert,

Eva Kondorosi………………………………………………………………………...147

Expression Pattern of DMI Genes in Medicago NodulesRossana Mirabella, Marijke Hartog, Carolien Franken, René Geurts and

Ton Bisseling…………………………………………………………………………..153

Actinorhizal Symbioses Katharina Pawlowski, Anna Zdyb, Bettina Hause, Cornelia Göbe, Ivo

Feussner, Kirill Demchenko…………………………………………………………..157

Signaling for Nodulation in a Water-Tolerant Legume Marcelle Holsters, Ward Capoen, Jeroen den Herder and Sofie Goormachtig………161

xv

Genetic and Molecular Analysis of Nod Factor Signalling in Medicago

truncatula

F. Debellé, C. Bres, J. Lévy, B. Ben Amor, J.F. Arrighi, F. Maillet, J.M. Ane,

C. Rosenberg, J. Dénarié, S. Shaw, G. Oldroyd, S. Long, R. Penmetsa, D.

Cook, R. Geurts, T. Bisseling, G. Duc and C. Gough…………………………………165

Activation and Perception of Calcium Oscillations during nod Factor SignallingCynthia Gleason, Raka Mitra, Péter Kaló, Christine Galera, Clare Gough,

Jean Dénarié, Sharon R. Long and Giles E.D. Oldroyd………………………………169

Functional Genomics of the Regulation of Nodule Number in Legumes Peter M. Gresshoff, Gustavo Gualtieri, Titeki Laniya, Arief Indrasumunar,

Akira Miyahara, Sureeporn Nontachaiyapoom, Tim Wells, Bandana Biswas,

Pick Kuen Chan, Paul Scott, M. Kinkema, M. Djordjevic, Dana Hoffmann,

Lisette Pregelj, Diana M. Buzas, Dong Xi Li, Artem Men, Qunyi Jiang,

Cheol-Ho Hwang and Bernard J. Carroll…………………………………………….173

“Activator” and “Inhibitor” Leading to Generation and Stabilization of Symbiotic Organ Development in Legume Masayoshi Kawaguchi………………………………………………………………...179

Regulatory Mechanisms of SYMRK Kinase Activity Satoko Yoshida and Martin Parniske………………………………………………….183

A Method to Enable Continuous Liquid Introduction into an Apoplast of Soybean-Plantlet Leaf and its Application to Elucidating the Supernodulation Trait of Nod1-3 Yasuhiro Arima, Hiroko Yamaya and Tadashi Yokoyama……………………………..187

Amino Acid Cycling by Rhizobium leguminosarum in Pea Nodules James White, Alex Bourdes, Arthur Hosie, Seonag Kinghorn, Philip Poole………….189

Root Nodule Extensins in Infection Thread Development Elizabeth A. Rathbun and Nicholas J. Brewin………………………………………...193

CASTOR and POLLUX, the Twin Genes that are Responsible for Endosymbioses in Lotus japonicus

Haruko Imaizumi-Anraku, Naoya Takeda, Martin Parniske, Makoto Hayashi,

Shinji Kawasaki……………………………………………………………………….195

The Expression of the Rhizobium tropici guaB Gene is Required in the Early Stages of Bean Nodulation Mónica Collavino, Daniel H. Grasso, Pablo M. Riccillo and O. Mario Aguilar……..199

xvi

Does the Rhizobial Type III Secretion System of Rhizobium Species NGR234 Affect the Defence Response in Plant Leaves? O. Schumpp, W.J. Deakin, C. Stahelin and W.J. Broughton………………………….201

Mathematical Modeling for Nodule Size Distribution Jun-ichi Ikeda, Ayako Fukunaga, Yuko Suga and Kaneaki Hori……………………..203

Early Events in the Nodulation of Casuarina glauca by Frankia

L. Laplaze, S. Svistoonoff, M. Obertello, B. Peret, F. Auguy, M.O. Sy,

V. Hocher, D. Autran, M. Nicole, C. Franche, D. Bogusz…………………………….205

Gene Expression in the Root Nodules of Elaeagnus umbellata

Chung-Sun An, Ho-Bang Kim, Sang-Ho Lee, Jung Jang Hyun, Chang-Jae

Oh, Hyoungseok Lee…………………………………………………………………..207

Diffusible Signal Factors in Nodulation of Discaria trinervis by Frankia

Luciano A. Gabbarini and Luis G. Wall………………………………………..……...209

Cloning and Expression Studies of furF and cap F in Frankia Strain R43 João Vieira, Pedro Moradas-Ferreira and Fernando Tavares………………………..211

Characterization of Mutants from Rhizobium sp. NGR234 with Defective Expolysaccharide Synthesis C. Staehelin, W. D’Haeze, M.Y. Gao, R.W. Carlson, B.J. Pellock, G.C. Walker,

W.R. Streit and W.J. Broughton………………………………………………………..213

Isolation of Two New Sinorhizobium meliloti Transcriptional Regulators Required for Nodulation Li Luo, Shi-Yi Yao, Anke Becker, Silvia Rüberg, Guan-Qiao Yu, Jia-Bi Zhu and

Hai-Ping Cheng……………………………………………………………………….215

Rhizobial Control of Host-Specificity N.M. Boukli, W.J. Deakin, K. Kambara, H. Kobayashi, C. Marie, X. Perret,

A. Le Quéeré, B. Reuhs, M. Saad, O. Schumpp, P. Skorpil, C. Staehelin,

W. Streit and W.J. Broughton…………………………………………………….……217

The Sinorhizobium melilot nifA Exerts its Signaling Effect for Nodulation in the Host Plants Huasong Zou, Guanqiao Yu and Jiabi Zhu……………………………………………219

xvii

Evolutionary Stability of Rhizobium Mutualism Depends on Legume Host SanctionsR. Ford Denison and E. Toby Kiers…………………………………………………...221

Poster Summaries……………………………………………………………………..225

SECTION V. TOWARDS SUSTAINABLE AGRICULTURE AND

PROTECTION OF THE ENVIRONMENT……………………………………….251

Ecological Significance of Lumichrome and Riboflavin as Signals in the Rhizosphere of Plants Felix D. Dakora, Sheku Kanu and Viviene N. Maitru...................................................253

Rhizospheric Plant-Microbe Interactions for Sustainable Agriculture Kauser A. Malik, F.Y. Hafeez, M.S. Mirza, S. Hameed, G. Rasul and

R. Bilal………………………………………………………………………………...257

Genetic Programs for Development of Nodules and Arbuscular Mycorrhiza in Legumes: Solid Facts and Unsolved Problems A.Y. Borisov, V.E. Tsyganov, A.O. Ovtsyna, N.A. Provorov, and I.A.

Tikhonovich……………………………………………………………………………261

Microbial Inoculants: A Challenge to Tune the Microbial Metabolites and Signals for Plant Responsiveness in the Field R. Remans, C. Snoeck, E. Luyten, S. Dobbelaere, E. Somers, A. Croonenborghs,

J. Michiels and J. Vanderleyden………………………………………………………265

Are Legumes Doing Their Job? The Effect of Herbicides on N2 Fixation in Southern Australian Agricultural Systems Elizabeth A. Drew, Vadakattu VSR Gupta, David K. Roget…………………………...269

Non-Symbiotic Bacterial Diazotrophs in Crop-Farming Systems: Can Their Potential for Plant Growth Promotion be Better Exploited? Ivan R. Kennedy, A.T.M.A. Choudhury, Mihály L. Keeskés, Rodney J. Roughley

and Nguyen Thanh Hien………………………………………………………………271

xviii

Effect of Legume Nodule Hydrogen Uptake Status on Hydrogen-Oxidizing Rhizobacteria and the Rotation of Crops Zhongmin Dong……………………………………………………………………….273

Phosphorous Use Efficiency for Symbiotic Nitrogen Fixation in Common Bean (Phaseolus vulgaris) and its Consequence on Soil P Dynamic Jean-Jacques Drevon, Nora Alkama, Mathew Blair, Aurelio Garcia, German

Hernandez, Philippe Hinsinger, Benoit Jaillard, Aline Lopez, Paula Rodino……….277

Mutational Analysis to Study the Role of Genetic Factors in Pea Adaptation to Stresses during Development its Symbioses with Rhizobium and Mycorrhizal FungiViktor E. Tsyganov, Alexander I. Zhernakov, Anna V. Khodorenko, Pavel Y.

Kisutin, Andrei A. Belimov, Vera I. Safronova, Tatyana S. Naumkina, Alexey

Y. Borisov, Peter Lindlbad, Karl-Josef Dietz, Igor A. Tikhonovich…………………..279

Effects of Waterlogging on Nitrogen Fixation of a Supernodulating Soybean Genotype, Sakukei 4 T. Matsunami, G.H. Jung, Y. Oki, W.H. Zhang and M. Kokubun……………………..283

Effect of N Application Rate on Nitrogen Fixation and Transfer from Vetch to Barley in Mixed Stand Hyowon Lee, Wongo Kim, Hyungsu Park and Sugon Kim…………………………...285

Analysis of the Promotive Effects of Deep Placement of Slow Release Fertilizers on Growth and Seed Yield of Soybean by 15N Dilution Method Tewari Kaushal, Masaru Onda, Sayuri Ito, Akihiko Yamazaki, Hiroyuki

Fujikake, Norikuni Ohtake, Kuni Sueyoshi, Yoshihiko Takahashi and

Takuji Ohayama………………………………………………………………………287

Role of Rhizobial Endophytes as Nitrogen Fixers in Promoting Plant Growth and Productivity of Indian Cultivated Upland Rice (Orysa sativa L.) Plants R.K. Singh, R.P.N. Mishra and H.K. Jaiswal…………………………………………289

Growth, Photosynthesis, Nodule Nitrogen and Carbon Fixation in Desi and Kabuli Genotypes of Chickpea (Cicer Arietinum L.) Under Salt StressGarg Neera and Ranju Singla………………………………………………………...293

Nodule Co-Occupancy of Agrobacterium and Bradyrhizobium with Potential Benefit to Legume Host Sohail Hameed, Fathia Mubeen, Kauser A. Malik and Fauzia Hafeez……………….295

xix

Salt Stress Adaptation in Alfalfa Bacteroids: Importance of Protein Betaine Karine Mandon, Alexandre Boscari, Jean Charles Trinchant, Laurence

Dupont, Geneviève Alloing, Didier Héouart, and Daniel Le Rudulier……………….297

Isolation of Genes for Salt Tolerance from Sinorhizobium LT11 W. Payakapong, P. Tittabutr, N. Teaumroong, D. Borthakur and N.

Boonkerd………………………………………………………………………………301

Regulation of Heme and Iron Metabolism in Bradyrhizobium japonicum

Jianhua Yang, Yali Friedman and Mark R. O’Brian………………………………….303

Rhizobial Genes Essential for Salt Tolerance Ju Quan Jiang, Wei Wei, Li Shi Xie, Lei Wang and Su Sheng Yang…………………...307

Effect of Low-pH and Aluminum on the Nodulation Signal Transduction in Medicago sativa

Min Yang, Xiaofeng Li, Yongxiong Yu and Minghua Gu………………………………309

Further Investigation of the Roles of Poly-3-Hydroxybutyrate (PHB) and Glycogen in Sinorhizobium meliloti Medicago sp. Symbiosis Chunxia Wang, Marsha Saldanha, Xiaoyan Sheng, Kris Shelswell,

Trevor C. Charles and Bruno W. Sorbral……………………………………………..311

Poster Summaries…………………………………………………………………..…313

SECTION VI. MOLECULAR ECOLOGY AND THE DISCOVERY

OF NEW NITROGEN FIXERS…………………………………………………….337

Diazotrophic Endophytes in Rice: Colonization and Nitrogen Fixation of Herbaspirillum and Clostridium Species Kiwamu Minamisawa, Mu You, Tadashi Abe, Bin Ye, Asami Saito, Makoto

Kawahara and Tadashi Sato…………………………………………………………..339

New Perspectives of Grass-Associated Nitrogen Fixation B. Reinhold-Hurek, F. Battistoni, A. Krause, A. Ramakumar, F. Friedrich,

A. Sarkar, M. Böhm, S. Gemmer, L. Miché, L. Zhang and T. Hurek…………………..345

Electron Donation and Alternative Nitrogenases in Cyanobacterial Dinitrogen FixationHermann Bothe and Gudrun Boison………………………………………………….349

xx

Role of Unicellular Diazotrophs in the Oceanic Nitrogen Fixation Joseph P. Montoya…………………………………………………………………….355

New Nitrogen-Fixing Microorganisms from the Oceans: Biological Aspects and Global Implications Jonathan P. Zehr, Barbara Methe and Rachel Foster………………………………...361

Biodiversity and Phylogeny of Rhizobial Germplasm in China Wen Xin Chen, En Tao Wang and Wen Feng Chen……………………………………367

The Species Paradigm in Bacteriology: From a Cross-Disciplinary Species Concept to a New Prokaryotic Species Definition (with Emphasis on Rhizobia)Kristina Lindström, Zewdu Terefework, Petri Auvinen, Lars Paulin, Stefan…………373Weidner, Anke Becker, Silvia Rüberg and Helge Gyllenberg

Direct Amplification of Rhizobial nodC Sequences from Soil Total DNA and Comparison to nodC Diversity of Root Nodule Isolates Sarita Sarita, Parveen K. Sharma, Ursula B. Priefer and Juergen Prell………….…377

Recent Studies on the Rhizobium-Cereal Association Frank B. Dazzo, Youssef G. Yanni, Rizk Rizk, M. Zidan, Abu-Bakr M.

Gomaa, Andrea Squartini, Yu-Xiang Jing, Feng Chi and Shi-Hua Shen……………..379

Ascending Migration of Endophytic Rhizobia from Roots to Leaves inside Rice Plants Feng Chi, Shi-Hua Shen, Hai-Ping Cheng, Yu-Xiang Jing and F.B. Dazzo………….381

The Ecology of Azorhizobium caulinodans ORS571 Colonizing on Root Surfaces of Arabidopsis thaliana

Taichiro Iki, Toshihiro Aono, Oyaizu Hiroshi…………………………………………383

Evolutionary Implication of Nitrogenase-Like Proteins in the Plant Kingdom and Prospects for nif Gene Transfer in Model Eukaryotes Q. Cheng, J. Yang, A. Day, M. Dowson-Day and R. Dixon…………………………...387

Novel Mimosa-Nodulating Strains of Burkholderia from South America Euan K. James, Wen-Ming Chen, Sergio M. De Faria, Jean L. Simões-Araujo,

Roseangela Straliotto, Rosa M. Pitard, Jui-Hsing Chou, Yi-Ju Chou,

Edmundo Barrios, Alan R. Prescott, Janet I. Sprent, J. Peter W. Young……………...391

xxi

Investigation Approach on Associative Diazotrophs in Plant Microecosystem with Multi-Field of View Song Wei………………………………………………………………………………395

Poster Summaries………………………………………………………….………….398

AUTHOR INDEX……………………………………………………………………431

xxiii

PREFACE

The 14th International Nitrogen Fixation Congress was held in Beijing, China from October 27th through November 1st, 2004. This volume constitutes the proceedings of the Congress and represents a compilation of the presentations by scientists from more than 30 countries around the World who came to Beijing to discuss the progress made since the last Congress and to exchange ideas and information. This year marked the 30th anniversary of the first Congress held in Pullman, Washington, USA, in 1974. Since then, this series of Congresses has met five times in North America (three in the United States and once each in Canada and Mexico), once in South America (Brazil), four times in Western Europe (once each in Spain, The Netherlands, Germany and France), once in Eastern Europe (Russia), and once in Australia; and now for the first time in Asia. China was a most appropriate choice because China is a big country with the largest population in the World, about 1.3 billion people, which is about 22% of the World’s population. It is traditionally an agricultural country, even though China has only 7% of the available farming land. This situation explains why agriculture and its productivity are major issues for the Chinese people, its government and the scientists in the field.

There are many serious problems facing us in the World today: the global population is growing rapidly and most of the people in poverty are in developing countries; the increase in the grain-production rate on a global scale continues to decrease (from 3% in the 1960’s to 2% in the 1980’s and 1990’s to an estimated 1.8% in 2010); and too much chemical fertilizer is being applied and causing even-more serious environmental problems. These continuing problems clearly indicate that the World needs to develop effective forms of sustainable agriculture that result in protection of our environment. These issues have led to increased attention, especially in Asia, Africa, and Latin America, to research on biological nitrogen fixation. As a result, China, for example, has launched highly effective research programs and provided significant support for all areas of nitrogen fixation. Therefore, to host this Congress was a very special occasion for the Chinese scientists who work in this area.

Spectacular progress has occurred in this field over the last 3 years and we were fortunate to have the major players in attendance at the Congress. The content of the final program involved extensive consultation with the members of the International Steering Committee and the International Program Advisory Committee. We thank them all for their very valuable input. Our program covers the most recent theories on how nitrogenase may have evolved, how the enzyme catalyzes substrate reduction and how it is regulated in response to molecular oxygen, fixed nitrogen, and light. It then considers the current advances in genomics, transcriptomics, and proteomics. Besides information on several rhizobium genomes whose sequencing was achieved some time ago, new information on the genome of endophytic bacteria, such as Herbaspirillum

and Azoarcus as well as that of Frankia, was discussed at the Congress. The machinery involved in establishing a nitrogen-fixing symbiotic relationship and its

xxiv

requirements for effective functioning is extremely complex. Recent information on the common pool of genes required for nodule formation in legumes, efficient mychorrization, and actinorhizal symbiosis as well progress in the signal-transduction pathways leading to the organogenesis of the nodule was presented.

A large part of this volume reflects the concern of having a productive agriculture that will respect the environment. In particular, the reader will find information on the physiological and molecular mechanisms that enable adaptation to high saline soils, selection of the legumes genotypes with high productivity or with the super-nodulation phenotype, together with data on bioremediation of harmful chemicals and preparation and formulation of bacterial inoculants. Molecular ecology is another subject of importance, since most of soil, rhizosphere and endophytic nitrogen-fixing organisms remain to be discovered. However, a very large number of new nitrogen-fixing species have been discovered recently, and an up-to-date description of the taxonomy of rhizobia as well as molecular tools to study bacteria-plant interactions are described in this volume.

We want to express our sincere thanks to all those who worked hard and long to help us in the organization of this Congress. We are especially grateful to those people who stayed with us when we were confronted by the unexpected SARS outbreak in Beijing in 2003, which cost us a one-year postponement of the Congress. We also thank all our scientific colleagues, in particular Profs. Shanjiong Shen, Jilun Li, and Qirui Cai for their understanding and support. We want to emphasize our gratitude for the financial support that we received from Peking University, National Laboratory of Protein Engineering and Plant Genetic Engineering, The National Key Fundamental Research Program of China (973 program), The National High Technology Research and Development Program of China (863 program), the National Natural Science Foundation of China, UNESCO, Sichuan Habio Bioengineering Co., Ltd and Shenfeng Biotech Engineering Co., Ltd of Angang Industry.

Finally, we all thank Vicki Newton for her extraordinarily efficient expertise during the editing of this proceedings volume. We also thank Beiyan Nan for his assistance with computing and related problems. Most importantly, we want to thank our families and friends for their moral support during the last three years and all the participants for making this Congress such an enjoyable and interesting experience for us. We hope you enjoyed it as much as we did.

Yi-Ping Wang Min Lin Zhe-Xian Tian Claudine Elmerich William E. Newton

Beijing, November, 2004

14TH INTERNATIONAL CONGRESS

ON NITROGEN FIXATION

DEDICATION

The editors dedicate this volume to

Jia-Xi Lu and

Hua-Kui Chen

in acknowledgement of their contributions to nitrogen fixation research in China

Professor Lu Jiaxi (1915-2001)