Polyphasic characterization of rhizobial isolates obtained from different common bean-producing regions
DOI:
https://doi.org/10.14393/BJ-v39n0a2023-67229Keywords:
Biological Nitrogen Fixation, Characterization, Diversity, Phaseolus vulgaris.Abstract
In Brazil, the common bean is a crop with significant social and economic importance. The prospecting of N2 fixing bacteria is crucial since biological nitrogen fixation (BNF) is an eco-friendly technique. This work aimed to obtain and characterize rhizobium isolates based on morpho-physiological, molecular, and symbiotic efficiency parameters, using the strains SEMIA 4077, SEMIA 4080, and SEMIA 4088 as references. The characteristics of the isolates and colonies, their tolerance to salinity and temperature, as well as their utilization of carbon sources, served as the basis for the morpho-physiological characterization. BOX-PCR, REP-PCR, and ERIC-PCR markers were used for genotypic characterization. Assessment of the symbiotic efficiency was carried out in a greenhouse, determining the number of nodules (NN), nodule dry weight (NDW), shoot dry weight (SDW), and total-N (Total-N) accumulation in the shoot. Among the isolates, those exhibiting: neutral culture medium pH, fast growth, colony diameter <2 mm, opaque transparency, homogeneous appearance, and cream color were predominant. Compared to temperature, salinity was the most restrictive factor to the growth of the isolates. Most of the isolates grew on sucrose (88.43%) and mannitol (87.28%). Genotypic analysis revealed that 90% of the isolates clustered in the same group as the reference strain SEMIA 4080. The TaMsG2R1 and BaDeG4R2 isolates showed higher Total-N in the shoot than the reference strains and should be evaluated in future studies under field conditions.
Downloads
References
ARGAW, A. and TSIGIE, A. Indigenous rhizobia population influences the effectiveness of Rhizobium inoculation and need of inorganic N for common bean (Phaseolus vulgaris L.) production in eastern Ethiopia. Chemical and Biological Technologies in Agriculture. 2015, 2(19), 2-13. https://doi.org/10.1186/s40538-015-0047-z
AUSUBEL, F.M. et al. Short Protocols in Molecular Biology. 4th ed. New York: John Wiley and Sons, Inc.: 1999.
CARDOSO, J.D., HUNGRIA, M. and ANDRADE, D.S. Polyphasic approach for the characterization of rhizobial symbionts effective in fixing N2 with common bean (Phaseolus vulgaris L.). Applied Genetics and Molecular Biotechnology. 2012, 93, 2035–2049. https://doi.org/10.1007/s00253-011-3708-2
CARDOSO, A.A. et al. Characterization of rhizobia isolates obtained from nodules of wild genotypes of common bean. Brazilian Journal of Microbiology. 2017, 48(1), 43-50. https://doi.org/10.1016/j.bjm.2016.09.002
CHUEIRE, L.M. et al. Taxonomic classification of rhizobial strains recommended for soybean and common bean crops in Brazil based on the sequencing of the 16s rRNA gene. Revista Brasileira de Ciência do Solo. 2013, 27, 833-840. https://doi.org/10.1590/S0100-06832003000500007
DALL’AGNOL, R.F. et al. Rhizobium freirei sp. nov., a symbiont of Phaseolus vulgaris that is very affective at fixing nitrogen. International Journal of Systemic and Evolutionary Microbiology. 2013, 63, 4167-4173. https://doi.org/10.1099/ijs.0.052928-0
EMBRAPA – Empresa Brasileira de Pesquisa Agropecuária. Manual de métodos de análise de solo. 2nd ed. Rio de Janeiro: Centro Nacional de Pesquisa de Solos, 1997. 212 p.
FERNANDES JÚNIOR, P.I. et al. Phenotypic diversity and amylolytic activity of fast growing rhizobia from pigeonpea [Cajanus cajan (L.) Millsp.]. Brazilian Journal of Microbiology. 2012, 43(4), 1604-1612. https://doi.org/10.1590/S1517-83822012000400045
FERREIRA, D.F. SISVAR: a computer analysis system to fixed effects splitplot type designs. Brazilian Journal of Biometrics. 2019, 37(4), 529–535. https://doi.org/10.28951/rbb.v37i4.450
HUNGRIA, M. 1994. Coleta de nódulos e isolamento de rizóbio. In: M. HUNGRIA, R.S. ARAÚJO, eds. Manual de métodos empregados em estudos de microbiologia agrícola. Santo Antônio de Goiás-GO, Embrapa Arroz e Feijão, pp. 45-61.
KASCHUK, G., HUNGRIA, M. and ANDRADE, D.S. Genetic diversity of rhizobia associated with common bean (Phaseolus vulgaris L.) grown under no-tillage and conventional systems in Southern Brazil. Applied Soil Ecology. 2006, 32(2), 210–220. https://doi.org/10.1016/j.apsoil.2005.06.008
KOSKEY, G. et al. Potential of Native Rhizobia in Enhancing Nitrogen Fixation and Yields of Climbing Beans (Phaseolus vulgaris L.) in Contrasting Environments of Eastern Kenya. Frontiers in Plant Science. 2017, 8. https://doi.org/10.3389/fpls.2017.00443
MARTINS, L.M.V. et al. Características relativas ao crescimento em meio de cultura e a morfologia de colônias de rizóbio. Seropédica-RJ. Embrapa Agrobiologia, 1997.
MILCHESKI, V.F. et al. Influência da interação entre genótipos de feijoeiro e rizóbios na nodulação e fixação de nitrogênio. Revista de Ciências Agroveterinárias. 2022, 21(1), 8-15. https://doi.org/10.5965/223811712112022008
MOREIRA, L.P., OLIVEIRA, A.P.S. and FERREIRA, E.P.B. Nodulation, contribution of biological N2 fixation, and productivity of the common bean (Phaseolus vulgaris L.) inoculated with rhizobia isolates. Australian Journal of Crop Science. 2017, 11(6), 644–651. https://doi.org/10.21475/ajcs.17.11.06.p310
NASCENTE, A.S. et al. Nitrogen management effects on soil mineral nitrogen, plant nutrition and yield of super early of super cycle common bean genotypes. Acta Scientiarum: Agronomy. 2017, 39, 369-378. https://doi.org/10.4025/actasciagron.v39i3.32781
PATEL, J. and DUBEY, S.K. Effect of different carbon source on growth of Rhizobium belonging to different region of Chhattisgarh, India. International Journal of Current Microbiology and Applied Sciences. 2014, 3(10), 830-836.
RAZIKA, G. et al. Influence of carbon source on the production of exopolysacharides by Rhizobium sullae and on the nodulation of Hedysarum coronarium L. legume. African Journal of Microbiology Research. 2012, 6(30), 5940-5946. https://doi.org/10.5897/AJMR12.393
ROHLF, F.J. NTSYS-pc: numerical taxonomy and multivariate analysis system. New York: Exeter Publication, 1988.
SAMPAIO, F.B., et al. Morphophysiological characterization of rhizobia isolated from wild genotypes of the common bean. Bioscience Journal. 2016, 32(6), 1502-1511. https://doi.org/10.14393/BJ-v32n6a2016-33084
SAMUDIN, S. and KUSWANTORO, H. Effect of Rhizobium inoculation to nodulation and growth of soybean [Glycine max (L.) Merrill] germplasm. Legume Research. 2018, 41(2), 303-310. https://doi.org/10.18805/LR-385
SANTIAGO-RAMOS, D. et al. Physicochemical properties of nixtamalized black bean (Phaseolus vulgaris L.) flours. Food Chemistry. 2018, 240, 456-462. https://doi.org/10.1016/j.foodchem.2017.07.156
SHAMSELDIN, A. and VELÁZQUEZ, E. The promiscuity of Phaseolus vulgaris L. (common bean) for nodulation with rhizobia: a review. World Journal of Microbiology and Biotechnology. 2020, 36(63). https://doi.org/10.1007/s11274-020-02839-w
SHARMA, S.R. et al. Isolation and characterization of salt-tolerant rhizobia native to the desert soils of United Arab Emirates. Emirates Journal of Food & Agriculture. 2013, 25(2), 102-108. https://doi.org/10.9755/ejfa.v25i2.7590
SILVA, D.J. and QUEIROZ, A.C. Análise de alimentos: métodos químicos e biológicos. 3rd ed. Viçosa: UFV, 2009. 235p.
SORATTO, R.P. et al. Nutrient extraction and exportation by common bean cultivars under different fertilization levels: I - macronutrients. Revista Brasileira de Ciência do Solo. 2013, 37(4), 1027-1042. https://doi.org/10.1590/S0100-06832013000400020
SOUZA, J.E.B. and FERREIRA, E.P.B. Improving sustainability of common bean production systems by co-inoculating rhizobia and azospirilla. Agriculture, Ecosystems and Environment, 2017, 237, 205-257. https://doi.org/10.1016/j.agee.2016.12.040
VERSALOVIC, J. et al. Genomic fingerprinting of bacteria using repetitive sequence-based polymerase chain reaction. Methods in molecular and cellular biology, 1994, 5(1), 25-40.
YANG, C. et al. Symbiosis of selected Rhizobium leguminosarum bv. viciae strains with diverse pea genotypes: effects on biological nitrogen fixation. Canadian Journal of Microbiology. 2017, 63(11), 909–919. https://doi.org/10.1139/cjm-2017-0281
YOUSEIF, S.H. et al. Phenotypic characteristics and genetic diversity of rhizobia nodulating soybean in Egyptian soils, European Journal of Soil Biology. 2014, 60, 34-43. https://doi.org/10.1016/j.ejsobi.2013.10.008
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2023 Thályta Duarte dos Santos, Princewill Chukwuma Asobia, Enderson Petrônio de Brito Ferreira
This work is licensed under a Creative Commons Attribution 4.0 International License.