Abstract
For over 7 centuries, production of rice (Oryza sativa L.) in Egypt has benefited from rotation with Egyptian berseem clover (Trifolium alexandrinum) The nitrogen supplied by this rotation replaces 25– 33% of the recommended rate of fertilizer-N application for rice production. This benefit to the rice cannot be explained solely by an increased availability of fixed N through mineralization of N- rich clover crop residues. Since rice normally supports a diverse microbial community of internal root colonists, we have examined the possibility that the clover symbiont, Rhizobium leguminosarum by. trifolii colonizes rice roots endophytically in fields where these crops are rotated, and if so, whether this novel plant-microbe association benefits rice growth. MPN plant infection studies were performed on macerates of surface-sterilized rice roots inoculated on T alexandrinum as the legume trap host. The results indicated that the root interior of rice grown in fields rotated with clover in the Nile Delta contained ~106 clover-nodulating rhizobial endophytes g−1 fresh weight of root. Plant tests plus microscopical, cultural, biochemical, and molecular structure studies indicated that the numerically dominant isolates of clover-nodulating rice endophytes represent 3 – 4 authentic strains of R. leguminosarum by. trifolii that were Nod+ Fix+ on berseem clover. Pure cultures of selected strains were able to colonize the interior of rice roots grown under gnotobiotic conditions. These rice endophytes were reisolated from surface-sterilized roots and shown by molecular methods to be the same as the original inoculant strains, thus verifying Koch’s postulates. Two endophytic strains of R leguminosarum by. trifolii significantly increased shoot and root growth of rice in growth chamber experiments, and grain yield plus agronomic fertilizer N-use efficiency of Giza-175 hybrid rice in a field inoculation experiment conducted in the Nile Delta. Thus, fields where rice has been grown in rotation with clover since antiquity contain Fix+ strains of R. leguminosarum by. trifolii that naturally colonize the rice root interior, and these true rhizobial endophytes have the potential to promote rice growth and productivity under laboratory and field conditions.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
Abbreviations
- GC/MS:
-
gas chromatography-mass spectrometry, 1
- H-NMR:
-
proton nuclear magnetic resonance spectroscopy,
- IRRI:
-
International Rice Research Institute,
- LSD:
-
least significant difference,
- MPN:
-
most probable number,
- PCR:
-
polymerase chain reaction,
- PGPR:
-
plant growth promoting rhizobacteria,
- RFLP:
-
restriction fragment length polymorphism,
- RDP:
-
ribosomal database project,
- SDS-PAGE:
-
sodium dodecylsulfatepolyacrylamide gel electrophoresis,
- YEM:
-
yeast extract mannitol
References
Abdel Wahab A M, Zahran H H, Abd-Alla M H 1996 Root-hair infection and nodulation of four grain legumes as affected by the form and the application time of nitrogen fertilizer. Folia Microbiol. 41, 303–308.
Balandreau J and Knowles R 1978 The rhizosphere. In Interactions Between Non-Pathogenic Soil Microorganisms and Plants. Eds. Y R Dommergues, S V Krupa. pp 243–268, Elsevier, Amsterdam.
Bally R, Thomas-Bauzon D, Heulin T, Balandreau J, Richard C and Ley J D 1983 Determination of the most frequent N2-fixing bacteria in a rice rhizosphere. Can. J. Microbiol. 29, 881–887.
Bilai R and Malik K A 1987 Isolation and identification of a N2- fixing zoogloea-forming bacterium from kallar grass histoplane. J. Appl. Bacteriol. 62, 289–294.
Black A D, Evans F, Ensminger J,White F, Clar J and Dinaver R 1965 Methods of Soil Analysis - II. Chemical and Microbiological Properties. No. 9 in the Series of Agronomy, American Society for Agronomy, Madison, WI.
Boddey R and Dobereiner J 1988 Nitrogen fixation associated with grasses and cereals: recent results and perspectives for future research. Plant Soil 108, 53–65.
Chabot R H, Antoun H, Kloepper J and Beauchamp C 1996 Root colonization of maize and lettuce by bioluminescent Rhizobium leguminosarum biovar phaseoli. Appl. Environ. Microbiol. 62, 2767–2772.
Cocking E C, Davey M R, Kothari S L, Srivastava J S, Jing Y, Ridge R W and Rolfe B G 1992 Altering the specificity control of the interaction between rhizobia and plants. Symbiosis 14, 123–130.
Conch V, Giacomini A, 011ero F J, Squartini A and Nuti M P 1991 Pulsed-field electrophoresis in contour-clamped homogeneous electric fields (CHEF) for the fingerprinting of Rhizobium spp. FEMS Microbiol. Lett. 83, 193–198.
Dazzo F B 1982 Leguminous root nodules. In Experimental Microbial Ecology. Ed. J Slater and R Burns. pp 431–446. Blackwell Scientific Publications, Oxford, UK.
Dazzo F B and Brill W J 1978 Regulation by fixed nitrogen of hostsymbiont recognition in the Rhizobium-clover symbiosis. Plant Physiol. 62, 18–21.
Dazzo F B, Orgambide G G, Philip-Hollingsworth S, Hollingsworth R I, Ninke K O and Salzwedel J L 1996 Modulation of development, growth dynamics, wall crystallinity, and infection sites in white clover root hairs by membrane chitolipooligosaccharides from Rhizobium leguminosarum biovar trifolii. J. Bacteriol. 178, 3621–3627.
Dazzo F B and Petersen J 1989 Applications of computer-assisted image analysis for microscopical studies of the Rhizobium-legume symbiosis. Symbiosis 7, 193–210.
DeBruijn F J,Jing Y and Dazzo F B 1995 Potentials and pitfalls of trying to extend symbiotic interactions of nitrogen-fixing organisms to presently non-nodulated plants, such as rice. Plant Soil 174,225–240.
DeSoete G 1984 Additive-tree representations of incomplete dissimilarity data. Quality Quantity 18, 387–393.
Dobereiner J, Reis V, Paula M and Olivares F 1993 Endophytic diazotrophs in sugar cane, cereals, and tuber plants. In New Horizons in Nitrogen Fixation. Eds. R Palacios, J Mora and W E Newton. pp 671–679. Kluwer Academic Publishers, Dordrecht, The Netherlands.
Eckhardt T 1978 A rapid method for the identification of plasmid deoxyribonucleic acid in bacteria. Plasmid 1, 584–588.
Espuny M R, 011ero F J, Bellogin R A, Ruiz-Sainz J E and Perez-Silva J 1987 Transfer of the Rhizobium leguminosarum biovar trifolii symbiotic plasmid pRtr5a to a strain of Rhizobium sp. that nodulates Hedysarum coronarium. J. Appl. Bacteriol. 63, 13–20.
Graves W L, Williams W A, Wegrzyn V A, Calderon D, George M R and Sullins J L 1987 Berseem clover is getting a second chance. California Agriculture September-October 1987, pp 15–18.
Gough C, Webster G, Vasse J, Galera C, Batchelor C, O’Callaghan K, Davey M, Denarie J and Cocking E 1996 Intercellular infection of wheat and Arabidopsis by rhizobial strains. Abstr. S-45, 8th International Congress on Molecular Plant-Microbe Interactions, Knoxville, TN.
Hashem F M and Kuykendall D 1994 Plasmid DNA content of several agronomically important Rhizobium species that nodulate alfalfa, berseem clover, or Leucaena. In Symbiotic Nitrogen Fixation. Eds. P Graham, M Sadowsky and C Vance. pp 181–188. Kluwer Academic Publishers, Dordrecht, The Netherlands.
Holflich G, Wiehe W and Hecht-Bucholz C 1995 Rhizosphere colonization of different crops with growth promoting Pseudomonas and Rhizobium bacteria. Microbiol. Res. 150, 139–147.
Hollingsworth R I, Squartini A, Philip-Hollingsworth S and Dazzo F B 1989 Root hair deforming and nodule initiating factors from Rhizobium trifolii. In Signal Molecules in Plants and Plant-Microbe Interactions. Ed. B Lugtenberg. pp 387–393. Springer-Verlag, Berlin, Germany.
Hurek T, Reinhold-Hurek B, van Montague M, and Kellenberger E 1994 Root colonization and systemic spreading of Azoarcus sp. strain BH72 in grasses. J. Bacteriol. 176, 1913–1923.
International Rice Research Institute 1996 IRRI towards 2020. International Rice Research Institute. P.O. Box 933, Manila, Philippines. 43 p.
Khush G S and Bennett J 1992 Nodulation and Nitrogen fixation in rice: Potential and prospects. International Rice Research Institute Press, Manila, Philippines. 136 p.
Klein D A, Salzwedel J L and Dazzo F B 1990 Microbial colonization of plant roots. In Biotechnology of Plant-Microbe Interactions. Eds. J P Nakas and C Hagedorn. pp 189–225. McGraw-Hill Publishing Company, NY.
Kloepper J W, Schippers B and Bakker P A 1992 Proposed elimina- tion of the term endorhizosphere. Phytopathol. 82, 726–727.
Ladha J K 1986 Studies on nitrogen fixation by free-living and rice-plant associated bacteria in wetland rice field. Bionature 6, 47–58.
Ladha J K, Tirol-Padre A, Reddy K and Ventura W 1993 Prospects and problems of biological nitrogen fixation in rice production: a critical assessment. In New Horizons in Nitrogen Fixation. Eds. R Palacios, J Mora and W E Newton. pp 677–682. Kluwer Academic Publishers, Dordrecht, The Netherlands.
Ladha J K, Garcia M, Miyan S, Padre A T and Watanabe I 1989 Survival of Azorhizobium caulinodans in the soil and rhizosphere of wetland rice under Sesbania rostrata-rice rotation. Appl. Environ. Microbiol. 55, 454–460.
Ladha J K, So R, Hernandez R, Dazzo F B, Reddy P M, Angeles O R, Ramos M C, de Bruijn F J and Stoltzfus J 1996a Rhizobial invasion and induction of phenotypic changes in rice roots are independent of Nod factors. Proceedings 8th International Congress on Molecular Plant-Microbe Interactions (Abstr. #L11), Knoxville, TN.
Laemmli U 1970 Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (London) 227, 680–685.
Larsen N, Olsen G, Maidak B, McCaughey M, Overbeek R, Macke T, Marsch T and Woese C R 1993 The ribosomal database project. Nucl. Acid Res. 21, 3021–3032.
Mateos P, Jiminez J, Chen J, Squartini A, Martinez-Molina E, Hubbell D H and Dazzo F B 1992 Cell-associated pectinolytic and cellulolytic enzymes in Rhizobium trifolii. Appl. Environ. Microbiol. 58, 1816–1822.
Natalia K, Gennady K and Vitaly K 1994 Endophytic association of nitrogen-fixing bacteria with plants. Proceedings 1st European N2-Fixation Conference (Abstr. P104 ), Szeged, Hungary.
Ndoye I, DeBilly F, Vasse J, Dreyfus B and Truchet G 1994 Root nodulation of Sesbania rostrada. J. Bacteriol. 176, 1060–1068.
Old K and Nicolson T 1975 Electron microscopical studies of the microflora of roots of sand dune grasses. New Phytol. 74, 51–58.
Old K and Nicolson T 1978 The root cortex as part of a microbial continuum. In Microbial Ecology. Eds. M Loutit and J Miles. pp 291–294. Springer-Verlag, NY.
Okon Y and Labandera-Gonzalez C A 1994 Agronomic applications of Azospirillum: an evaluation of 20 years worldwide field inoculation. Soil Biol. Biochem. 26, 1591–1601.
Orgambide G, Lee J, Hollingsworth R and Dazzo F B 1995 Structurally diverse chitolipooligosaccharide Nod factors accumulate primarily in membranes of wild type Rhizobium leguminosarum by. trifolii. Biochemistry 34, 3832–3840.
Orgambide G, Philip-Hollingsworth S, Hollingsworth R I and Dazzo F B 1994 Flavone-enhanced accumulation and symbiosis-related biological activity of a diglycosyl diacylglycerol membrane glycolipid from Rhizobium leguminosarum by. trifolii. J. Bacteriol. 176, 4338–4347.
Orgambide G, Philip-Hollingsworth S, Mateos P F, Hollingsworth R I and Dazzo F B 1996 Subnanomolar concentrations of membrane chitolipooligosaccharides from Rhizobium leguminosarum by. trifolii are fully capable of eliciting symbiosis-related responses on white clover. Plant Soil, 186, 93–98.
Philip-Hollingsworth S, Hollingsworth R I and Dazzo F B 1989 Host-range related structural features of the acidic extracellular polysaccharides of Rhizobium trifolii and Rhizobium leguminosarum. J. Biol. Chem. 264, 1461–1466.
Philip-Hollingsworth S, Hollingsworth R I and Dazzo F B 1991 N-acetylglutamic acid: an extracellular Nod signal of Rhizobium trifolii ANU843 which induces root hair branching and nodule-like primordia in white clover roots. J. Biol. Chem. 266, 16854–16858.
Reddy P M, Ladha J K, So R, Hernandez R, Dazzo F B, Angeles O R, Ramos M C and de Bruijn F J 1997 Rhizobial communication with rice: induction of phenotypic changes, mode of invasion and extent of colonization. Plant Soil 194, 81–98.
Roger P and Ladha J K 1992 Biological nitrogen fixation in wetland rice fields: estimation and contribution to nitrogen balance. Plant Soil 141, 41–55.
Roger P A and Watanabe I W 1986 Technologies for utilizing biological nitrogen fixation in wetland rice: potentialities, current usage, and limiting factors. Fert. Res. 9, 39–77.
Shimshick E J and Herbert R R 1979 Binding characteristics of N2-fixing bacteria to cereal roots. Appl. Environ. Microbiol. 38, 447–453.
Smucker A J 1993 Soil environmental modifications of root dynam- ics and measurement. Annu. Rev. Phytopathol. 31, 191–216.
Somasegaran P and Holben H J 1985 Methods in Legume-Rhizobium Technology. NifI’AL Project, University of Hawaii, Maui, Hawaii. 367 p.
Spencer D, James E K, Ellis G J, Shaw J E and Sprent J I 1994 Interaction between rhizobia and potato tissues. J. Exp. Bot. 45, 1475–1482.
Subba-Rao N S, Mateos P F, Baker D, Pankratz H S, Palma J, Dazzo F B and Sprent J I 1995 The unique root-nodule symbiosis between Rhizobium and the aquatic legume, Neptunia moans (L. f.) Druce. Planta 196, 311–320.
Tien T, Gaskins M H and Hubbell D H 1979 Plant growth substances produced by Azospirillum brasilense and their effect on the growth of pearl millet (Pennisetum americanum L.). Appl. Environ. Microbiol. 37, 1016–1024.
Ueda T, Suga Y, Yahiro N and Matsuguchi T 1995 Remarkable N2-fixing bacterial diversity detected in rice roots by molecular evolutionary analysis of nifH gene sequences. J. Bacteriol. 177, 1414–1417.
Umali-Garcia M, Hubbell D H, Gaskins M H and Dazzo F B 1980 Association of Azospirillum with grass roots. Appl. Environ. Microbiol. 39, 219–226.
Versalovic J, Schneider M, de Bruijn F J and Lupski J R 1994 Genomic fingerprinting of bacteria using repetitive sequence-based polymerase chain reaction. Methods Molec Cell. Biol 5, 25–40.
Willems A and Collins M D 1993 Phylogenetic analysis of rhizobia and agrobacteria based on 16S rRNA gene sequences. Int. J. Syst. Bacteriol. 43, 305–313.
Yanni Y G 1991 Potential of indigenous cyanobacteria to contribute to rice performance under different schedules of nitrogen application. World J. Microbiol. Biotech. 7, 48–52.
Young J P and Haukka K E 1996 Diversity and phylogeny of rhizobia. New Phytol. 133, 87–94.
Author information
Authors and Affiliations
Corresponding author
Editor information
Rights and permissions
Copyright information
© 1997 Springer Science+Business Media Dordrecht
About this chapter
Cite this chapter
Yanni, Y.G. et al. (1997). Natural endophytic association between Rhizobium leguminosarum bv. trifolii and rice roots and assessment of its potential to promote rice growth. In: Ladha, J.K., de Bruijn, F.J., Malik, K.A. (eds) Opportunities for Biological Nitrogen Fixation in Rice and Other Non-Legumes. Developments in Plant and Soil Sciences, vol 75. Springer, Dordrecht. https://doi.org/10.1007/978-94-011-7113-7_10
Download citation
DOI: https://doi.org/10.1007/978-94-011-7113-7_10
Publisher Name: Springer, Dordrecht
Print ISBN: 978-0-7923-4748-4
Online ISBN: 978-94-011-7113-7
eBook Packages: Springer Book Archive