FRANKIA AND ACTINORHIZAL PLANTS

Developments in Plant and Soil Sciences Volume 18

I. 1. Monteith and C. Webb, eds., Soil Water and Nitrogen in Mediterranean-type Environments. 1981. ISBN 90-247-2406-6

2. 1.C. Brogan, ed., Nitrogen Losses and Surface Run-off from Landspreading of Manures. 1981. ISBN 90-247-2471-6

3. 1.D. Bewley, ed., Nitrogen and Carbon Metabolism. 1981. ISBN 90-247-2472-4

4. R. Brouwer, l. Gasparikova, 1. Kolek and B.C. Loughman, eds., Structure and Function of Plant Roots. 1981. ISBN 90-247-2510-0

5. Y.R. Dommergues and H.G. Diem, eds., Microbiology of Tropical Soils and Plant Productivity. 1982. ISBN 90-247-2624-7

6. G.P. Robertson, R. Herrera and T. Rosswall, eds., Nitrogen Cycling in Ecosystems of Latin America and the Caribbean. 1982. ISBN 90-247-2719-7

7. D. Atkinson et aI., eds., Tree Root Systems and their Mycorrhizas. 1983. ISBN 90-247-2821-5

8. M.R. Saric and B.C. Loughman, eds., Genetic Aspects of Plant Nutrition. 1983. ISBN 90-247-2822-3

9. 1.R. Freney and 1.R. Simpson, eds., Gaseous Loss of Nitrogen from Plant-Soil Systems. 1983. ISBN 90-247-2820-7

10. United Nations Economic Commission for Europe. Efficient Use of Fertilizers in Agriculture. 1983. ISBN 90-247-2866-5

11. 1. Tinsley and 1.F. Darbyshire, eds., Biological Processes and Soil Fertility. 1984. ISBN 90-247-2902-5

12. A.D.L. Akkermans, D. Baker, K. Huss-Danell and 1.D. Tjepkema, eds., Frankia Symbioses. 1984. ISBN 90-247-2967-X

13. W.S. Silver and E.C. Schroder, eds., Practical Application of Azalla for Rice Production. 1984. ISBN 90-247-3068-6

14. P.G.L. Vlek, ed., Micronutrients in Tropical Food Crop Production. 1985. ISBN 90-247-3085-6

15. T.P. Hignett, ed., Fertilizer Manual. 1985. ISBN 90-247-3122-4

16. D. Vaughan and R.E. Malcolm, eds., Soil Organic Matter and Biological Activity. 1985. ISBN 90-247-3154-2

17. D. Pasternak and A. San Pietro, eds., Biosalinity in Action: Bioproduction with Saline Water 1985. ISBN 90-247-3159-3. In Press

18. M. Lalonde, C. Camire and 1.0. Dawson, eds., Frankia and Actinorhizal Plants. 1985. ISBN 90-247-3214-X

Frankia and Actinorhizal Plants

Edited by

M. LALONDE, c. CAMIRE Departement des sciences jorestieres Faculte de joresterie Universite Laval Ste-Foy, Que. Canada, G1K 7P4

and

J.O. DAWSON Forestry Department University oj Illinois Urbana, IL 61801, USA

First published as Plant and Soil, Vol. 87, No.1 (1985)

1985 MARTINUS NIJHOFF PUBLISHERS .. a member of the KLUWER ACADEMIC PUBLISHERS GROUP 1Ii11 DORDRECHT / BOSTON / LANCASTER •

Proceedings of the International Symposium on Frankia and Actinorhizal Plants, held in Montmorency Forest, Laval University, Quebec, Canada on 5-9 August 1984

Distributors

jor the United States and Canada: Kluwer Academic Publishers, 190 Old Derby Street, Hingham, MA 02043, USA jor the UK and Ireland: Kluwer Academic Publishers, MTP Press Limited, Falcon House, Queen Square, Lancaster LAI lRN, UK jor all other countries: Kluwer Academic Publishers Group, Distribution Center, P.O. Box 322, 3300 AH Dordrecht, The Netherlands

Library of Congress Catalogue Card Number: 85-15366

ISBN-13: 978-94-010-8777-3 DOl: 10 .1007/978-94-009-514 7 -1

Copyright

e-ISBN-13: 978-94-009-5147-1

© 1985 by Martinus Nijhoff Publishers, Dordrecht. Softcover reprint of the hardcover 1 st edition 1985

All rights reserved. No part of this publication may be reproduced, stored in a retrieval system, or transmitted in any form or by any means, mechanical, photocopying, recording, or otherwise, without the prior written permission of the publishers, Martinus Nijhoff Publishers, P.O. Box 163, 3300 AD Dordrecht, The Netherlands.

Contents

Registrants

Preface

Z. Zhongze and 1.G. Torrey, Studies of an effective strain of Frankia from Allocasuarina lehmanniana of the Casuarinaceae

H.G. Diem and Y.R. Dommergues, In vitro production of specialized reproductive torulose hyphae by Frankia strain ORS 021001 isolated

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from Casuarinajunghuhniana root nodules 17

W.L. Parson, L.R. Robertson and C.V. Carpenter, Characterization and infectivity of a spontaneous variant isolated from Frankia sp. WEY 0131391 31

C.S. An, W. S. Riggsby and B.C. Mullin, Restriction pattern analysis of genomic DNA of Frankia isolates 43

P. Simonet, P. Normand, A. Moiroud and M. Lalonde, Restriction enzyme digestion patterns of Frankia plasmids 49

H. liabin, Z. Zheying, C. Guanxiong and L. Huichang, Host range of Frankia endophytes 61

L. Vergnaud, A. Chaboud, Y. Prin and M. Rougier, Preinfection events in the establishment of Alnus-Frankia symbiosis: Development of a spot inoculation technique 67

C.S. Vogel and 1.0. Dawson, Effect of juglone on growth in vitro of Frankia isolates and nodulation of Alnus glutinosa in soil 79

L.l. Winship and 1.D. Tjepkema, Nitrogen fixation and respiration by root nodules of Alnus rubra Bong.: Effects of temperature and oxygen concentration

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C.R. Schwintzer, Effect of spring flooding on endophyte differentia-tion, nitrogenase activity, root growth and shoot growth in Myrica gale 109

L. Simon, A. Stein, S. Cote and M. Lalonde, Performance of in vitro propagated Alnus glutinosa (L.) Gaertn. clones inoculated with Frankiae 125

T.C. Hennesey, L.K. Bair and R.W. McNew, Variation in response among three Alnus spp. clones to progressive water stress 135

L.J. Bertrand and M. Lalonde, In vitro propagation and nodulation by Frankia of actinorhizal Russian Olive (Elaeagnus angustifolia L.) 143

M.D. Stowers and J.E. Smith, Inoculation and production of container-grown red alder seedlings 153

A.M. Berry and J .G. Torrey, Seed germination, seedling inoculation and establishment of Alnus spp. in containers in greenhouse trials 161

P. Perinet, J.G. Brouillette, J.A. Fortin and M. Lalonde, Large scale inoculation of actinorhizal plants with Frankia 175

G. Pregent and C. Camire, Biomass production by alders on four abandoned agricultural soils in Quebec 185

B. Cote and C. Camire, Nitrogen cycling in dense plantings of hybrid poplar and black alder 195

Registrants

A. C. Aldrich, Dept. of Microbiology and Cell Science, 1059 McCarty Hall, IFAS, University of Florida, Gainesville, FL 32611, USA

C. An, Dept. of Botany, University of Tennessee, Knoxville, TN 37916, USA

J. A. Arnonne, Forestry and Environmental Studies, 370 Prospect St., New Haven, CT 06511, USA

D. Bair, Forestry Dept., Iowa State University, Ames, IA 50013, USA

D. Baker, CF Kettering Research Lab., 150 E. South College, Yellow Springs, OH 45387, USA

D. Benson, Dept. of Biochemistry and Microbiology, Cook College, Rutgers Univ., New Bruns­wick, NJ 08903, USA

H. R. Berg, Dept. of Microbiology and Cell Science, 1059 McCarty Hall, IF AS, University of Florida, Gainesville, FL 32611, USA

A. M. Berry, Dept. of Environmental Horticulture, University of California, Davis, CA 95616, USA

L. J. Bertrand, Dep. des sciences forestieres, Fac. de foresterie et de geodesie, Universite Laval, Quebec, Canada, GIK 7P4

1. Bousquet, Dep. des sciences forestieres, Fac. de foresterie et de geodesie, Universite Laval, Quebec, Canada G 1 K 7P4

J. G. Brouillette, Societe d'energie de la Baie James, 800 Bou!. de Maisonneuve est Montreal, Quebec, Canada, H2L 4MB

D. A. Callaham, Botany Department, University of Massachusetts, Amherst, MA 01003, USA

C. Camire, Dep. des sciences forestieres, Fac. de foresterie et de geodesie, Universite Laval, Quebec, Canada, GIK 7P4

C. V. Carpenter, Weyerhaeuser Company, WTC 2B25, Tacoma, WA 98477, USA

A. Chaboud, Dep. de biologie vegetale, Universite de Lyon I, 43 Boulevard du 11 Novembre 1918, F-69622 Villeurbanne, France

Guanxiong, Chen, Institute of Forestry and Soil Science, Academia Sinica, Shenyang, China

J. Collins, Dept. of Botany and Microbiology, Oklahoma State University, Stillwater, OK 74078, USA

B. Cote, Dept. of Forestry, University of Illinois, Urbana, IL 61801, USA

S. Cote, Dep. des sciences forestieres, Fac. de foresterie et de geodesie, Universite Laval, Quebec, Canada, GIK 7P4

J. O. Dawson, Dept. of Forestry, University of Illinois, Urbana, IL 61801, USA

H. G. Diem, ORSTOM/CNRS, B. P. 1386, Dakar, Senegal

Y. Dommergues, ORSTOM/CNRS, B. P. 1386, Dakar, Senegal

J. C. Ensign, Dept. of Bacteriology, University of Wisconsin, Madison, WI 53706, USA

M. S. Fontaine, Cabot Foundation, Harvard University, Petersham, MA 01366, USA

J. A. Fortin, Dep. des sciences forestieres, Fac. de foresterie et de geodesie, Universite Laval, Quebec, Canada, GIK 7P4

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M. Gardes, Dep. des sciences forestieres, Fac. de foresterie et de geodesie, Universite Lava!, Quebec, Canada, GIK 7P4

G. Z. Gertner, Dept. of Forestry, University ofIJIinois, Urbana, IL 61801, USA

1. Giasson, Dep. des sciences forestieres, Fac. de foresterie et de geodesie, Universite Laval, Quebec, Canada, GIK 7P4

T. C. Hennessey, Dept. of Forestry, Oklahoma State University, Stillwater, OK 74074, USA

P. K. Hepler, Botany Department, University of Massachusetts, Amherst, MA 01003, USA

A. Hirsch, Dept. of Biological Sciences, Wellesley College, Wellesley, MA 02181, USA

A. HQuwers, Plant and Soil, Lab. Microbiology, H. van Suchtelenweg 4, 6703 CTWageningen, The Netherlands

L. Huichang, Institute of Forestry and Soil Science, Academia Sinica, Shenyang, China

D. Idie, University of Tennessee, Knoxville, TN 37996, USA

S. Jagar, Dept. of Botany, University of Tennessee, Knoxville, TN 37916, USA

H. Jiabin, Institute of Forestry and Soil Science, Academic Sinica, Shenyang, China

S. Laliberte, Dep. des sciences forestil~res, Fac. de foresterie et de geodesie, Universite Laval, Quebec, Canada, GIK 7P4

M. Lalonde, Dep. des sciences forestieres, Fac. de foresterie et de geodesie, Universite Laval, Quebec, Canada, GIK 7P4

S. Lancelle, Cabot Foundation, Harvard University, Petersham, MA 01366, USA

H. A. Lechevalier, Waksman Institute of Microbiology, Rutgers University, P. O. Box 759, Piscataway, NJ 08854, USA

M. P. Lechevalier, Waksman Institute of Microbiollgy, Rutgers University, P. O. Box 759, Piscataway, NJ 08854, USA

J. Ligan, Allied Corporation, P. O. Box 6, Solvay, NY 13209, USA

M. F. Lopez, Cabot Foundation, Harvard University, Petersham, MA 01366, USA

E. M. Lorenzi, Dept. of Forestry, Oklahoma State University, Stillwater, OK 74074, USA

H. Massicotte, Botany and Genetics, University of Guelph, Guelph, Ont., Canada, NIG 2Wl

M. E. McCully, Biology Department, Carleton University, Ottawa, Ont., Canada, KIS 5B6

1. McDowell, Dept. of Microbiology and Cell Science, 1059 McCarty Hall, IFAS, University of Florida, Gainesville, FL 32611, USA

1. McIntyre, Biology Department, Carleton University, Ottawa, Ont., Canada, KIS 5B6

C. Miguel, Unit of Nitrogen Fixation, Centro de Edafologia y Biologia Aplicada, C.S.I.C., Apartado 257, Salamanca, Spain

A. Moiroud, Universite de Lyon I, Boulevard du 11 Novembre 1918, F-69622 Villeurbanne, France

B. C. Mullin, Dept. of Botany, University of Tennessee, Knoxville, TN 37996, USA

A. Nesme, Biofutur/INRA, F-49000 Angers, France

P. Normand, Dep. des sciences forestieres, Fac. de foresterie et de geodesie, Universite Lava!, Quebec, Canada, GIK 7P4

W. Parson, Lab. 11-2249, Evergreen State College, Olympia, WA 98505, USA

M. Pelchat, Dep. des sciences forestieres, Fac. de foresterie et de geodesie, Universite Laval, Quebec, Canada, GIK 7P4

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P. Perinet, Rhizotec Laboratories Inc., C. P. 797, St-Jean-Chrysostome, Quebec, Canada, G6Z 2L9

Y. Piche, Petawawa National Forestry Institute, Chalk River, Ont., Canada, KOJ 110

G. Pregent, Dep. des sciences forestj(~res, Fac. de foresterie et de geodesie, Universite Laval, Quebec, Canada, GIK 7P4

Y. Prin, Dep. de Biologie vegetale, Universite de Lyon I, 43 Boulevard du 11 Novembre 1918, F-69622 Villeurbanne, France

e. Rioux, Dept. of Microbiology, University of Guelph, Guelph, Ont., Canada, NIG 2Wl

M. Roberts, Dept. of Botany, University of Tennessee, Knoxville, TN 37916, USA

L. R .. Robertson, Weyerhaeuser Company, WTC 2B25, Tacoma, WA 98477, USA

e. Rodriguez-Barrueco, Unit of Nitrogen Fixation, Centro de Edafologia y Biologia Aplicada, C.S.I.e., Apartado 257, Salamanca, Spain

M. Rougier, Dep. de Biologie Vegetale, Universite de Lyon I, 43 Boulevard du 11 Novembre 1918, F-69622 Villeurbanne, France

C. R. Schwintzer, Dept. of Botany and Plant Pathology, University of Maine, Orono, ME 04469, USA

L. Simon, Dep. des sciences forestieres, Fac. de foresterie et de geoctesie, Universite Laval, Quebec, Canada, GIK 7P4

P. Simonet, Dep. des sciences forestieres, Fac. de foresterie et de geodesie, Universite Laval, Quebec, Canada, GIK 7P4

J. E. Smith, NPI, University Research Park, Salt Lake City, UT 84108, USA

A. Stein, Dep. des sciences forestieres, Fac. de foresterie et de geodesie, Universite Laval, Quebec, Canada, GIK 7P4

L. St-Laurent, Dep. des sciences forestieres, Fac. de foresterie et de geodesie, Universite Laval, Quebec, Canada, GIK 7P4

M. D. Stowers, NPI, University Research Park, Salt Lake City, UT, 84108, USA

P. Subramaniam, Unit of Nitrogen Fixation, Centro de Edafologia y Biologia Aplicada, C.S.I.C., Apartado 257, Salamanca, Spain

L. S. Tisa, Dept. of Bacteriology, University of Wisconsin, Madison, WI 53706, USA

J. Tjepkema, Dept. of Botany and Plant Pathology, University of Maine, Orono, ME 04469, USA

J. G. Torrey, Cabot Foundation, Harvard University, Petersham, MA 01366, USA

F. Tremblay, Dep. des sciences forestieres, Fac. de foresterie et de geodesie, Universite Laval, Quebec, Canada, GIK 7P4

M. F. Tremblay, Dep. des sciences forestieres, Fac. de foresterie et de geodesie, Universite Laval, Quebec, Canada, GIK 7P4

S. Tremblay, Dep. des sciences forestieres, Fac. de foresterie et de geoctesie, Universite Laval, Quebec, Canada, GIK 7P4

L. Vergnaud, Dep. de biologie vegetale, Universite de Lyon I, 43 Boulevard du 11 Novembre 1918, F-69622 Villeurbanne, France

H. S. Vishniac, Dept. of Botany and Microbiology, Oklahoma State University, Stillwater, OK 74078, USA

C. S. Vogel, Dept. of Forestry, University of Illinois, Urbana, IL 61801, USA

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A. Weber, Dept. of General Microbiology, Mannerheimintie 172, SF-00280, Helsinki 28, Suomi/Finland

L. J. Winship, School of Natural Science, Hampshire College, Amherst, MA 01002, USA

P. Younger, Dep. des sciences forestieres, Fac. de foresterie et de geoctesie, Universite Laval, Quebec, Canada, G1K 7P4

Zhongze, Zhang, Cabot Foundation, Harvard University, Petersham, MA 01366, USA

Zhenying, Zhao, Institute of Forestry, Academia Sinica, Shenyang, China

Preface

The fifth meeting of scientists working with Frankia and actinorhizal plants was held at Montmorency Forest of Laval University in Quebec from August 6-8, 1984. Results of research presented at the meeting are included in this special volume of Plant and Soil. The understanding and use of actinorhizal systems continue to increase, though work­shops and discussions at this and similar meetings make it evident that this important subject remains open for fruitful investigation at all levels.

Some important 'firsts' were reported at this meeting. The first extensive survey of Frankiae and their host specificity ranges from Asia was presented. This is of significance since Asia is a center of diversity for many actinorhizal host plant genera. A report that proto­plasts of Frankia have been produced and regenerated for the first time improves the possibility for genetic manipulation of Frankia. It is also important to note the first report herein of successful mass inoculation of actinorhizal plants commercially for stabilization and reclamation of disturbed soils around hydroelectric power projects in Quebec. This heralds the transfer of actinorhizal technology to private and public users.

The bacterial genus Frankia is easily recognized both in vivo and in vitro, and isolation of this organism has become routine. But, as yet, there are not sufficient biochemical, morphological, or anatomical criteria for establishing species.

Although isolation of Frankia has become routine for many actino­rhizal associations, there are no isolates reported for about half of the known actinorhizal genera and some Frankia isolates cannot be manipu­lated so as to successfully reinfect the host plant from which they were obtained.

The analysis of DNA from Frankia and its host plants, and the genetic analysis of Frankia plasm ids, are emerging as tools for under­standing and manipulating actinorhizal symbioses. There is a need to find useful markers for distinguishing functions of genetic material from Frankia and its host plants. Investigations of metabolism are yielding pathways and barriers for biochemical transformations of Frankia.

Improved methods for researching actinorhizal partners reported at this meeting include freeze substitution of Frankia for electron micros­copy, the use of microbeads for precision placement of Frankia colon­ies near root surfaces, and tissue culture of host plants for studies of interactions between host and endophyte.

Unusual spore-like structures similar to chlamydospores were re­ported to occur on an isolate from Casuarina nodules at this meeting. Vesicles of Frankia have been purified and Frankia protoplasts have

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been produced for improved studies of organismal functions. Explor­ation of the events, preceeding and associated with the infection process continues, with extensive serial sectioning and ultrastructural examination of actinorhizal associates. Isolates from diverse geographic sources are now available, and cross-inoculation patterns continue to emerge.

Physiological and ecological studies of actinorhizal organisms are revealing relationships among respiration, growth, nitrogenase activity, nodulation, temperature, oxygen tension, water stress, mineral nutri­tion, allelochemicals, and other properties of actinorhizal organisms and their environment. Methods to optimize operations for inoculation of actinorhizal plants are also being developed.

Results of studies of pure and mixed plantations of Alnus and Populus trees illustrate the complexities of nitrogen fixation and move­ment in actinorhizal ecosystems. The effects of planting mixture, planting density, soil type, site, time, different actinorhizal plants, interplanted species, allelochemicals, weather, and other factors are important in determining plantation performance.

Future research needs include more Frankia isolations for taxonomic and practical use. It is also possible that there are more actinorhizal plants to be found, since Frankia has proven to be a very promiscuous nitrogen-fixing symbiont. Detailed study of the infection process is also needed. Such studies may lead to increased host susceptibility and the spread of symbiotic nitrogen fixation capabilities to additional host plant taxons. Recent advances in the understanding of manipu­lation of microbial genes have been spectacular. More emphasis and effort are needed in order to realize similar progress with Frankia. Much needs to be done in order to obtain a clearer picture of the phys­iology and biochemistry of Frankia and its host plants.

The need for information on actinorhizal systems seems to be particularly critical in developing countries where demand for fuel­wood, improved soil nitrogen fertility, and building materials can be better met with more effective use of actinorhizal plants. Furthermore, the Ubiquitous occurrence of actinorhizal plants in temperate eco­systems where they are often overlooked, suggest that research is needed to quantify current benefits so that we can better appreciate and improve our use of actinorhizal plants.

Jeffrey O. Dawson Forestry Department University of Illinois Urbana, Illinois, USA

Claude Camire and Maurice Lalonde Departement des sciences forestieres

Faculte de Foresterie Universite Laval

Quebec, Que. Canada