Register      Login
Soil Research Soil Research Society
Soil, land care and environmental research
RESEARCH ARTICLE

Medicago ciliaris growing in Tunisian soils is preferentially nodulated by Sinorhizobium medicae

K. Zribi A C , Y. Badri A , S. Saidi A , P. van Berkum B and M. E. Aouani A
+ Author Affiliations
- Author Affiliations

A Laboratoire Interactions Légumineuses Microorganismes (LILM), Centre de Biotechnologie, Technopole de Borj Cedria, BP901, Hammam lif 2050, Tunis, Tunisie.

B SGIL, Bldg-006, BARC-West, ARS, U.S. Department of Agriculture, 10300 Baltimore Blvd, Beltsville, MD 20705, USA.

C Corresponding author. Email: kais.zribi@cbbc.rnrt.tn

Australian Journal of Soil Research 45(6) 473-477 https://doi.org/10.1071/SR07083
Submitted: 28 February 2007  Accepted: 28 August 2007   Published: 20 September 2007

Abstract

Variation in growth of Medicago ciliaris was recorded across soils from 5 different regions in Tunisia that represented different soil types and climatic zones. In 4 of these soils (Mateur, Enfidha, Rhayet, and Soliman) this variation appeared to be related to the nodule number on the roots of the plants. With the exception of one isolate the rhizobia isolated from these nodules had 16S rRNA PCR-RFLP fingerprint patterns that were characteristic of Sinorhizobium medicae. Plant growth in the fifth soil (Jelma) was the poorest; plants had few nodules that yielded exclusively rhizobia with 16S rRNA fingerprint patterns characteristic of S. meliloti. In subsequent plant tests, S. medicae isolates formed effective nitrogen fixation symbioses with M. ciliaris, while S. meliloti formed small, white, ineffective nodules. Therefore, plant growth in Jelma soil was poor because only S. meliloti are present and this species is ineffective with M. ciliaris. In a co-inoculation experiment with M. ciliaris, S. medicae was more competitive for nodulation than S. meliloti, perhaps explaining why the majority of the isolates from Enfidha and Rhayet were S. medicae, since S. meliloti is present in these soils. However, it is not clear how the host influences rhizobia for nodulation by S. medicae in preference to S. meliloti when present.

Additional keywords: Medicago ciliaris, Sinorhizobium meliloti, Sinorhizobium medicae, PCR/RFLP, REP-PCR, nodule occupancy, symbiosis.


Acknowledgments

The authors thank M. Badri for providing seeds of Medicago ciliaris and A. Abdelguerfi and M. Laouar for helpful discussion.


References


Abdelly C, Lachaal M, Grignon C, Soltani A, Hajji M (1995) Association épisodique d’halophytes stricts et de glycophytes dans un écosystème hydromorphe salé en zone semi-aride (Episodic association of strict halophytes and glycophytes in a saline, hydromorphic ecosystem in semi-arid zones). Agronomie 15, 557–568.
Crossref | GoogleScholarGoogle Scholar | open url image1

Andronov EE, Roumyantseva ML, Sagoulenko VV, Simarov BV (1999) Effect of the host plant on the genetic diversity of a natural population of Sinorhizobium meliloti. Russian Journal of Genetics 35, 1169–1176. open url image1

Aouani ME, Mhamdi R, Jebara M, Amarger N (2001) Characterization of rhizobia nodulating chickpea in Tunisia. Agronomie 21, 577–581.
Crossref | GoogleScholarGoogle Scholar | open url image1

Badri Y, Zribi K, Badri M, Huguet T, Aouani ME (2003) Sinorhizobium meliloti nodulates Medicago laciniata in Tunisian soils. Czech Journal of Genetics & Plant Breeding 39, 178–183. open url image1

Bailly X, Olivieri I, De Mita S, Cleyet-Marel JC, Béna G (2006) Recombination and selection shape the molecular diversity pattern of nitrogen-fixing Sinorhizobium sp. associated to Medicago. Molecular Ecology 15, 2719–2734.
PubMed |
open url image1

Barran LR, Bromfield ESP, Brown DCW (2002) Identification and cloning of the bacterial nodulation specificity gene in the Sinorhizobium melilotiMedicago laciniata symbiosis. Canadian Journal of Microbiology 48, 765–771.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Ben Amor B, Shaw SL, Oldroyd GED, Maillet F, Penmetsa RV, Cook D, Long SR, Dénarié J, Gough C (2003) The NFP locus of Medicago truncatula controls an early step of Nod factor signal transduction upstream of a rapid calcium flux and root hair deformation. The Plant Journal 34, 495–506.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Béna G, Lyet A, Huguet T, Olivieri I (2005) MedicagoSinorhizobium symbiotic specificity evolution and the geographic expansion of Medicago. Journal of Evolutionary Biology 18, 1547–1559.
PubMed |
open url image1

van Berkum P, Elia P, Eardly BD (2006) Multilocus sequence typing as an approach for population analysis of Medicago-nodulating rhizobia. Journal of Bacteriology 188, 5570–5577.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

de Billy F, Grosjean C, May S, Bennett M, Cullimore JV (2001) Expression studies on Aux1-like genes in Medicago truncatula suggest that auxin is required at two steps in early nodule development. Molecular Plant-Microbe Interactions 14, 267–277.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Biondi EG, Pilli E, Giuntini E, Roumiantseva ML, Andronov EE , et al. (2003) Genetic relationship of Sinorhizobium meliloti and Sinorhizobium medicae strains isolated from Caucasian region. FEMS Microbiology Letters 220, 207–213.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Bromfield ESP, Barran LR, Wheatcroft R (1995) Relative genetic structure of population of Rhizobium meliloti isolated directly from soil and from nodules of alfalfa (Medicago sativa) and sweet clover (Melilotus alba). Molecular Ecology 4, 183–188. open url image1

Brunel B, Rome S, Ziani R, Cleyet-Marel JC (1996) Comparison of nucleotide diversity and symbiotic properties of Rhizobium meliloti populations from annual Medicago species. FEMS Microbiology Ecology 19, 71–82.
Crossref | GoogleScholarGoogle Scholar | open url image1

Carelli M, Gnocchi S, Fancelli S, Mengoni A, Paffetti D, Scotti C, Bazzicalupo M (2000) Genetic diversity and dynamics of Sinorhizobium meliloti populations nodulating different alfalfa cultivars in Italian soils. Applied and Environmental Microbiology 66, 4785–4789.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Cook D (1999) Medicago truncatula: a model in the making! Current Opinion in Plant Biology 2, 301–304.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

De Bruijn FJ (1992) Use of repetitive sequences and the polymerase chain reaction to fingerprint the genomes of Rhizobium meliloti isolates and other soil bacteria. Applied and Environmental Microbiology 58, 2180–2187.
PubMed |
open url image1

Fahråeus G (1957) The infection of clover root hairs by nodule bacteria studied by a simple glass slide technique. Journal of General Microbiology 16, 374–381.
PubMed |
open url image1

Garau G, Reeve WG, Brau L, Deiana P, Yates RJ, James D, Tiwari R, O’Hara GW, Howieson JG (2005) The symbiotic requirements of different Medicago spp. suggest the evolution of Sinorhizobium meliloti and S. medicae with hosts differentially adapted to soil pH. Plant and Soil 276, 263–277.
Crossref | GoogleScholarGoogle Scholar | open url image1

Hartmann A, Giraud JJ, Catroux G (1998) Genotypic diversity of Sinorhizobium (formerly Rhizobium) meliloti strains isolated directly from a soil and from nodules of alfalfa (Medicago sativa) grown in the same soil. FEMS Microbiology Ecology 25, 107–116.
Crossref | GoogleScholarGoogle Scholar | open url image1

Irwin JAG, Lloyd DL, Lowe KF (2001) Lucerne biology and genetic improvement—An analysis of past activities and future goals in Australia. Australian Journal of Agricultural Research 52, 699–712.
Crossref | GoogleScholarGoogle Scholar | open url image1

Jamann S, Fernandez MP, Normand P (1993) Typing method for N2-fixing bacteria based on PCR/RFLP- application to the characterization of Frankia strains. Molecular Ecology 2, 17–26.
PubMed |
open url image1

Jebara M, Mhamdi R, Aouani ME, Ghrir R, Mars M (2001) Genetic diversity of Sinorhizobium populations recovered from different Medicago varieties cultivated in Tunisian soils. Canadian Journal of Microbiology 47, 139–147.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Laguerre G, van Berkum P, Amarger N, Prevost D (1997) Genetic diversity of rhizobial symbionts isolated from legume species within the genera Astragalus, Oxutropis, and Onobrychis. Applied and Environmental Microbiology 63, 4748–4758.
PubMed |
open url image1

Laguerre G, Nour SM, Macheret V, Sanjuan J, Drouin P, Amarger N (2001) Classification of rhizobia based on nodC and nifH gene analysis reveals a close phylogenetic relationship among Phaseolus vulgaris symbionts. Microbiology 147, 981–993.
PubMed |
open url image1

Laouar M, Abdelguerfi A (2000) Etude du complexe d’espèces Medicago ciliaris-Medicago intertexta: caractérisations des différents types d’infrutescences. Cahiers Options Méditerranéennes 45, 39–41. open url image1

Lesins KA , Lesins I (1979) ‘Genus Medicago (Leguminosae): a taxogenetic study.’ pp. 46–53. (W. Junk Publishers: The Hague)

Mendes IC, Bottomley PJ (1998) Distribution of a population of Rhizobium leguminosarum bv. trifolii among different size classes of soil aggregates. Applied and Environmental Microbiology 64, 970–975. open url image1

Paffetti D, Daguin F, Fancelli S, Gnocchi S, Lippi F, Scotti C, Bazzicalupo M (1998) Influence of plant genotype on the selection of nodulating Sinorhizobium meliloti by Medicago sativa. Antonie van Leeuwenhoek 73, 3–8.
Crossref | GoogleScholarGoogle Scholar | open url image1

Paffetti D, Scotti C, Gnocchi S, Fancelli S, Bazzicalupo M (1996) Genetic diversity of an Italian Rhizobium meliloti population from Medicago sativa varieties. Applied and Environmental Microbiology 62, 2279–2285. open url image1

Rome S, Brunel B, Normand P, Fernandez MP, Cleyet-Marel JC (1996) Evidence that two genomic species of Rhizobium are associated with Medicago truncatula. Archives of Microbiology 165, 285–288.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Roumiantseva ML, Andronov EE, Sharypova LA, Dammann-Kalinowski T, Keller M, Young JPW, Simarov BV (2002) Diversity of Sinorhizobium meliloti from the Central Asian alfalfa gene center. Applied and Environmental Microbiology 68, 4694–4697.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Villegas MC, Rome S, Mauré L, Domergue O, Gardan L, Bailly X, Cleyet-Marel JC, Brunel B (2006) Nitrogen-fixing sinorhizobia with Medicago laciniata constitute a novel biovar (bv. medicaginis) of S. meliloti. Systematic and Applied Microbiology 29, 526–538.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Vincent JM (1970) ‘A manual for practical study of root-nodule bacteria.’ IBP handbook 15. (Blackwell Scientific Publications: Oxford, UK)

Zahran HH (1999) Rhizobium–Legume symbiosis and nitrogen fixation under severe conditions and in an arid climate. American Society for Microbiology (Ed.). Microbiology and Molecular Biology Reviews 63, 968–989.

Zribi K, Mhamdi R, Huguet T, Aouani ME (2004) Distribution and genetic diversity of rhizobia nodulating natural populations of Medicago truncatula in Tunisian soils. Soil Biology & Biochemistry 36, 903–908.
Crossref | GoogleScholarGoogle Scholar | open url image1

Zribi K, Mhamdi R, Huguet T, Aouani ME (2005) Diversity of Sinorhizobium meliloti and S. medicae nodulating Medicago truncatula according to host and soil origins. World Journal of Microbiology & Biotechnology 21, 1009–1015.
Crossref | GoogleScholarGoogle Scholar | open url image1