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Characterisation and evaluation ofRhizobium leguminosarum bv.trifolii nodulating rhizobia isolated from white clover native to Azad Jammu and Kashmir, Pakistan

  • Ecological and Environmental Microbiology
  • Original Articles
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Abstract

The present study was conducted to evaluate and characteriseRhizobium leguminosarum bv.trifolii nodulating rhizobia isolated from root nodules of indigenous and exotic ecotypes of white clover (Trifolium repens L.) in the mountain region of the State of Azad Jammu and Kashmir Pakistan. Isolates from indigenous ecotypes were collected from three different locations i.e. Tollipir, Banjosa, and Rawalakot while exotic ecotypes i.e. Irrigation and NuSiral were collected from Australia. Preliminary characterisation of isolates was carried-out on the basis of colony characteristics, phosphorus solubilisation, osmotic potential, relative absorbance, indole acetic acid spot test, mean doubling time and C-source utilisation.Rhizobium population in the respective soils was also estimated by using most probable number (MPN). The maximumRhizobium population of 5.8 × 106 g−1 soil was recorded in Tollipir soil compared with population of 3.1. × 105 g−1 soil in Rawalakot soil. All strains showed homology to a typicalRhizobium strain and identified asRhizobium on the basis of growth pattern and other testing parameters. The isolates were Gram negative, small rods and acid producing fast growers with a mean generation time in between 2.7 to 3.4 h. All isolates except isolates from Tollipir showed resistant against four antibiotics except tetracycline. Isolates from Banjosa, Tollipir and Rawalakot (indigenous) had larger surface charge than isolates from Irrigation and NuSiral (exotic). Highest osmotic potential of −31.02 bar was recorded in the isolates from Banjosa while isolates from Tollipir and Rawalakot had an osmotic potential of −22 bar. Field evaluation of isolated isolates showed significant increase in plant growth and root nodulation. The study will be useful for identifying the isolates under laboratory conditions and developing successful inoculation strategy for the production of white clover in the mountain region of Azad Jammu and Kashmir.

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References

  • Abbasi M.K., Tahir M.M., Khizar A. (2008). Forage production, nitrogen fixation and soil N accumulation of white clover (Trifolium repens L.) in the hill farming system of Azad Jammu and Kashmir. Commun. Soil Sci. Plant Anal. (in Press).

  • Ashraf M., Hassan F., Chaudhry I.A. (2002). Interaction ofRhizobium strain in lentil genotypes under rain fed conditions. Pak. J. Agric., Agric. Eng. Vet. Sci., 18: 38–43.

    Google Scholar 

  • Baryosef B., Rogers R.D., Wolfram J.W. (1999).Pseudomonas cepacia mediated rock phosphate solubilization in kaolinite and montmorillonite suspension. Soil Sci. Soc. Am. J., 63: 1703–1708.

    CAS  Google Scholar 

  • Ben Romdhane S., Nasar H., Samba-Mbaye R., Neyra M., Ghobral M.H. (2005). Genetic diversity ofAcacia tortilis rhizobia revealed by PCR/RFLP on crushed nodules in Tunisia. Ann. Microbiol., 55: 249–258.

    Google Scholar 

  • Ben Romdhane S., Elarbi Aquani M., Mhamdi R. (2007). Inefficient nodulation of chickpea (Cicer aeritinum L.) in the arid and Saharan climates in Tunisia bySinorhizobium meliloti biovarmedicaginis. Ann. Microbiol., 57: 15–19.

    Article  Google Scholar 

  • Bremner J.M., Mulvaney C.S. (1982). Nitrogen-total. In: Page A.L., Miller, R.H., Keeney D.R., Eds, Methods of Soil Analysis, Part 2, Chemical and Microbiological Properties, American Society of Agronomy, Madison, WI, USA, pp. 595–624.

    Google Scholar 

  • Burton J.C. (1985).Rhizobium relationships. In: Taylor N.I., Ed., Clover Science and Technology. (Agronomy Monograph no. 25). Soil Science Society of America, Madison, WI, USA.

    Google Scholar 

  • Bushby H.V.A. (1990). The role of bacterial surface charge in the ecology of root nodule bacteria. A hypothesis. Soil Biol. Biochem., 22: 1–9.

    Article  Google Scholar 

  • Busse M., Bottomley P. (1989). Growth and Nodulation responses ofRhizobium meliloti to water stress induced by permeating and non-permeating solutes. J. Appl. Environ. Microbiol., 55: 2431–2436.

    CAS  Google Scholar 

  • Chen W., Huang Q., Xiong X. (2004). Distribution and biodiversity of soybean rhizobia in the soils of Shennongjia forest reserve, China. Biol. Fertil. Soils., 40: 306–312.

    Article  CAS  Google Scholar 

  • Danso S.K.A., Owiredu J.D. (1988). Competitiveness of introduced and indigenous cowpeaBradyrhizobium strains for nodule formation in cowpeas in three soils. Soil Biol. Biochem., 20: 305–310.

    Article  Google Scholar 

  • Dravid M.S. (1991). Effect of salinization,Rhizobium nodulation, genotype variations and phosphorus application on dry matter yield and utilization of phosphorus by pea and lentil. J. Nuc. Agri. Biol., 9: 1227–1231.

    Google Scholar 

  • Duncan D.B. (1995). Multiple-range and multipleF-test. Biometrics. 11: 1–42.

    Article  Google Scholar 

  • Friederick J.B., Hagedorn C., Vanscyoc S.W. (1990). Isolation ofRhizobium leguminosarum (biovartrifolii). strains from Ethiopian soils and symbiotic effectiveness on African annual clover species. J. Appl. Environ. Microbiol., 56: 1087–1092.

    Google Scholar 

  • Gannon J.T., Manilal V.B., Alexander M. (1991). Relationship between cell surface properties and transport of bacteria through soil. J. Appl. Environ. Microbiol., 57: 190–193.

    CAS  Google Scholar 

  • Gordon S.A., Weber R.P. (1951). Colorimetric estimation of indole acetic acid. Plant Physiol., 26: 192–195.

    Article  CAS  PubMed  Google Scholar 

  • Haahtela K., Konkko R., Laakso T., Williams P.H., Korhonen T.N. (1990). Root associatedEnterobacter andKlebsiella inPoa pratensis: Characterization of an iron scaveging system and a substance stimulating root hair production. Mol. Plant Microb. Interact., 3: 358–365.

    CAS  Google Scholar 

  • Hafeez F.Y., Asad S., Ahmad T., Malik K.A. (1995). Host specificity and characterization of fast growing rhizobia fromMacroptilium atropurpureum cv. Siratro in Pakistan. Soil Biol. Biochem., 27: 729–733.

    Article  CAS  Google Scholar 

  • Harrower K.M., Nagy L.A. (1979). Effects of nutrients and water stress on growth and sorpulation of caprophilous fungi. Trans. British Mycol. Soc., 72: 459–462.

    Article  CAS  Google Scholar 

  • Hoagland D.R., Arnon D.I. (1950). The water culture method for growing plants without soil. California Berkeley Agriculture Circle. No. 347.

  • Kokhar S.N., Khan M.A., Chaudhri M.F. (2001). Some characters of chickpea-nodulating rhizobia native to Thal soil. Pak. J. Biol. Sci., 4: 1016–1019.

    Article  Google Scholar 

  • Kucey R.M.N. (1989). Response of field bean to levels ofRhizobium inoculation in soils containing effectiveleguminosarum phaseoli population. Can. J. Plant. Sci., 69: 419–426.

    Google Scholar 

  • Kuklinsky-Sobral J., Araujo W.L., Mendas R., Geraldi I.O., Pizzirani-Kleiner A.A., Azevedo J.L. (2004). Isolation and characterization of soybean-associated bacteria and their potential for plant growth promotion. Environ. Microbiol., 6: 1244–1251.

    Article  CAS  PubMed  Google Scholar 

  • Mnasri B., Mrabet M., Laguerre G., Aouani M.E., Mhamdi R. (2006). Salt-tolerant rhizobia from a Tunisian oasis that are highlt effective for N2-fixationwith Phaseoulus vulgaris constitute a novel biovar (bv.mediterranense) ofSinorhizobium meliloti. Arch. Microbiol., 187: 79–85.

    Article  PubMed  CAS  Google Scholar 

  • Nelson L.M., Child J.J. (1981). Nitrogen fixation and hydrogen metabolism byRhizobium leguminosarum isolates in pea root nodules. Can. J. Biol. Sci., 27: 1028–1034.

    CAS  Google Scholar 

  • Okon Y., Albercht S.L., Burris R.H. (1977). Methods for growingSpirillum lipoferum and for counting it in pure culture and in association with plants. Appl. Environ. Microbiol., 33: 85–88.

    PubMed  CAS  Google Scholar 

  • Pikovskaia R.L. (1948). Metabolisation of phosphorus in soil in connection with vital activity of some microbial species. Microbiology, 17: 362–370.

    Google Scholar 

  • Pryor H.N., Lowther W.L., Melntyre H.J., Ronson C.W. (1998).An inoculantRhizobium strain for improved establishment and growth of hexapliod Causasian Clover. New Zeal. J. Agric. Res., 41: 179–189.

    Google Scholar 

  • Rennie R.J. (1981). A single medium for the isolation of nitrogen fixing bacteria. Can. J. Microbiol., 27: 8–14.

    Article  CAS  PubMed  Google Scholar 

  • Rupela O.P., Beck D.F. (1990). Prospects for optimizing biological nitrogen fixation in Chickpea In: Rheenen and Saxena, Eds, Chickpea in the Nineties. Proceedings of the Second International Workshop on Chickpea Improvement, 4–8 Dec, 1989. ICRISAT.

  • Serraj R., Sinclair T. R. (1998). Soybean cultivar variability for nodule formation and growth under drought. Plant Soil, 202: 159–166.

    Article  CAS  Google Scholar 

  • Somasegaran P., Hoben H. (1985). Legume-Rhizobium Technology Manual. NifTAL Hawaii, USA.

    Google Scholar 

  • Tan I.K.P., Broughton W.I. (1981). Rhizobia in tropical legumes, X111, biochemical basis of acid and alkali reactions. Soil Biol. Biochem., 13: 389–393.

    Article  CAS  Google Scholar 

  • Tesfaye M., Holl F.B. (1999).Rhizobium strains that nodulateTrifolium semipilosum Fres. are phylogenetically distinct. Plant Soil, 207: 147–154.

    Article  Google Scholar 

  • Thies J.E., Singleton P.W., Bohool B.B. (1991). Influence of the size of the indigenous rhizobial populations on establishment and symbiotic performance of introduced rhizobia on field grown legumes. Appl. Environ. Micribiol., 57: 19–28.

    CAS  Google Scholar 

  • Vincent J.M. (1970). A Manual for the Practical Study of Root-Nodule Bacteria, IBP Handbook no. 15, Blackwell, Oxford.

    Google Scholar 

  • Yasmin S., Bakar M.A.R., Malik K.A., Hafeez F.Y. (2004). Isolation, characterization and beneficial effects of rice associated plant growth promoting bacteria from Zanzibar soils. J. Basic Microbiol., 44: 241–252.

    Article  CAS  PubMed  Google Scholar 

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Authors and Affiliations

  1. Department of Soil and Environmental Sciences, University of Azad Jammu and Kashmir, Faculty of Agriculture, Rawalakot, Azad Jammu and Kashmir (AJK), Pakistan

    Majid Mahmood Tahir & M. Kaleem Abbasi

  2. National Institute for Biotechnology and Genetic Engineering (NIBGE), Jhang Road, P.O. Box 577, 3800, Faisalabad, Pakistan

    Fauzia Y. Hafeez

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  1. Majid Mahmood Tahir
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  2. M. Kaleem Abbasi
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  3. Fauzia Y. Hafeez
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Correspondence to M. Kaleem Abbasi.

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Tahir, M.M., Abbasi, M.K. & Hafeez, F.Y. Characterisation and evaluation ofRhizobium leguminosarum bv.trifolii nodulating rhizobia isolated from white clover native to Azad Jammu and Kashmir, Pakistan. Ann. Microbiol. 58, 181–188 (2008). https://doi.org/10.1007/BF03175314

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  • DOI: https://doi.org/10.1007/BF03175314

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