Abstract
The application of commercial rhizobial inoculants to legume crops is proving to be an alternative to synthetic fertilizer use. The challenge for sustainable agriculture resides in the compatibility between crop, inoculants and environmental conditions. The evaluation of symbiotic efficiency and genetic diversity of indigenous rhizobial strains could lead to the development of better inoculants and increased crop production. The genetic variability of 32 wild indigenous rhizobial isolates was assessed by RAPD (Random Amplified Polymorphic DNA). The strains were isolated from red clover (Trifolium pratense L.) nodules from two distinct geographical regions of Northern and Eastern Romania. Three decamer primers were used to resolve the phylogenetic relationships between the investigated isolates. Cluster analysis revealed a high diversity; most strains clustered together based on their geographical location.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Castro-Sowinski, S., Herschkovitz, Y., Okon, Y., Jurkevitch, E. (2007) Effects of inoculation with plant growth-promoting rhizobacteria on resident rhizosphere microorganisms. FEMS Microbiol. Lett. 276, 1–11.
De Meyer, S. E., Van Hoorde, K., Vekeman, B., Braeckman, T., Willems, A. (2011) Genetic diversity of rhizobia associated with indigenous legumes in different regions of Flanders (Belgium). Soil Biol. Biochem. 43, 2384–2396.
De Oliveira, I. A., Vasconcellos, M. J., Seldin, L., Paiva, E., Vargas, M. A., de Sá, N. M. H. (2000) Random amplified polymorphic DNA analysis of effective Rhizobium sp. associated with beans cultivated in brazilian cerrado soils. Braz. J. Microbiol. 31, 39–44.
Durán, D., Rey, L., Sánchez-Cañizares, C., Navarro, A., Imperial, J., Ruiz-Argueso, T. (2013) Genetic diversity of indigenous rhizobial symbionts of the Lupinus mariae-josephae endemism from alkalinelimed soils within its area of distribution in Eastern Spain. Syst. Appl. Microbiol. 36, 128–136.
Giongo, A., Ambrosini, A., Vargas, L. K., Freire, J. R. J., Bodanese-Zanettini, M. H., Passaglia, L. M. P. (2008) Evaluation of genetic diversity of bradyrhizobia strains nodulating soybean [Glycine max (L.) Merrill] isolated from South Brazilian fields. Appl. Soil Ecol. 38, 261–269.
Hampl, V., Pavlícek, A., Flegr, J. (2001) Construction and bootstrap analysis of DNA fngerprintingbased phylogenetic trees with the freeware program FreeTree: application to trichomonad parasites. Int. J. Syst. Evol. Micr. 51, 731–735.
Herridge, D. F., Peoples, M. B., Boddey, R. M. (2008) Global inputs of biological nitrogen fixation in agricultural systems. Plant Soil 311, 1–18.
Iqbal, M. A., Khalid, M., Shahzad, S. M., Ahmad, M., Soleman, N., Akhtar, N. (2012) Integrated use of Rhizobium leguminosarum, plant growth promoting rhizobacteria and enriched compost for improving growth, nodulation and yield of lentil (Lens culinaris Medik.). Chil. J. Agr. Res. 72, 104–110.
Joseph, B., Ranjan Patra, R., Lawrence, R. (2012) Characterization of plant growth promoting rhizobacteria associated with chickpea (Cicer arietinum L.). Int. J. Plant Prod. 1, 141–152.
Karthikeyan, K. A. M., Sasireka, G., Suresh, P. (2013) Genetic diversity of Rhizobium leguminosarum isolates as revealed by Random Amplified polymorphic DNA analysis. J. Biosci. Res. 4, 15–20.
Lindström, K., Murwira, M., Willems, A., Altier, N. (2010) The biodiversity of beneficial microbehost mutualism: the case of rhizobia. Res. Microbiol. 161, 453–463.
Moschetti, G., Peluso, A., Protopapa, A., Anastasio, M., Pepe, O., Defez, R. (2005) Use of nodulation pattern, stress tolerance, nodC gene amplification, RAPD-PCR and RFLP-16S rDNA analysis to discriminate genotypes of Rhizobium leguminosarum biovar viciae. Sys. Appl. Microbiol. 28, 619–631.
Op den Camp, R. H., Polone, E., Fedorova, E., Roelofsen, W., Squartini, A., Op den Camp, H. J., Bisseling, T., Geurts, R. (2012) Nonlegume Parasponia andersonii deploys a broad rhizobium host range strategy resulting in largely variable symbiotic effectiveness. Mol. Plant Microbe In. 25, 954–963.
Paffetti, D., Scotti, C., Gnocchi, S., Fancelli, S., Bazzicalupo, M. (1996) Genetic diversity of an Italian Rhizobium meliloti population from different Medicago sativa varieties. Appl. Environ. Microb. 62, 2279–2285.
Pavel, A. B., Vasile, C. I. (2012) PyElph - a software tool for gel images analysis and phylogenetics. BMC Bioinformatics 13, 9.
Pinto, P. P., Paiva, E., Purcino, H., Passos, R. V. M., de Sá, N. M. H. (2004) Characterization of rhizobia that nodulate Arachis pintoi by RAPD analysis. Braz. J. Microbiol. 35, 219–223.
Rajasundari, K., Ilamurugu, K., Logeshwaran, P. (2009) Genetic diversity in rhizobial isolates determined by RAPDs. Afr. J. Biotechnol. 8, 2677–2681.
Ruiz-Díez, B., Quiñones, M. A., Fajardo, S., López, M. A., Higueras, P., Fernández-Pascual, M. (2012) Mercury-resistant rhizobial bacteria isolated from nodules of leguminous plants growing in high Hg-contaminated soils. Appl. Microbiol. Biotechnol. 96, 543–554.
Schluter, P. M., Harris, S. A. (2006) Analysis of multilocus fingerprinting data sets containing missing data. Mol. Ecol. Notes 6, 569–572.
Sikora, S., Redžepovic, S., Pejic, I., Kozumplik, V. (1997) Genetic diversity of Bradyrhizobium japonicum field population revealed by RAPD fingerprinting. J. Appl. Microbiol. 82, 527–531.
Sokal, R. R., Michener, C. D. (1958) A statistical method for evaluating systematic relationships. Univ. Kans. Sci. Bull. 28, 1409–1438.
Suominen, L., Jussila, M. M., Mäkeläinen, K., Romantschuk, M., Lindström, K. (2000) Evaluation of the Galega-Rhizobium galegae system for the bioremediation of oil-contaminated soil. Environ. Pollut. 107, 239–244.
Torres, A. R., Cursino, L., Muro-Abad, J. I., Gomes, E. A., de Araújo, E. F., Hungria, M., Cassini, S. T. A. (2009) Genetic diversity of indigenous common bean (Phaseolus vulgaris L.) rhizobia from the state of Minas Gerais, Brazil. Braz. J. Microbiol. 40, 852–856.
Vincent, J. M. (1970) A Manual for the Practical Study of Root-nodule Bacteria. IBP Handbook no. 15. Blackwell Scientific Publications, Oxford, England.
Wani, P. A., Khan, M. S. (2012) Bioremediation of lead by a plant growth promoting Rhizobium species RL9. Bacteriol. J. 2, 66–78.
Williams, J. G., Kubelik, A. R., Livak, K. J., Rafalski, J. A., Tingey, S. V. (1990) DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res. 18, 6531–6535.
Young, C. C., Cheng, K. T. (1998) Genetic diversity of fast-and slow-growing soybean rhizobia determined by random amplified polymorphic DNA analysis. Biol. Fert. Soils 26, 254–256.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
About this article
Cite this article
Stefan, A., Rosu, C.M., Stedel, C. et al. Rapd-Inferred Genetic Variability of Some Indigenous Rhizobium leguminosarum Isolates from Red Clover (Trifolium pratense L.) Nodules. BIOLOGIA FUTURA 66, 316–325 (2015). https://doi.org/10.1556/018.66.2015.3.7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1556/018.66.2015.3.7