Environmental Monitoring and Assessment

, Volume 184, Issue 1, pp 515–526 | Cite as

Isolation and characterization of metal resistant-tolerant rhizosphere bacteria from the serpentine soils in Turkey



Despite the number of studies describing metal hyper-accumulating plants and their associated bacteria in various regions and countries, there is no information on rhizosphere microbial potential of the Turkish serpentine soils. This study aimed to explore the rhizosphere microbial diversity of Ni-resistant, hyper-accumulating plants grown on Ni-rich soils and their metal tolerance–resistance characteristics. One hundred ninety-one locations were visited to collect soil and plant samples from different serpentine regions of Western Turkey. Following bioavailable and total Ni analysis of collected samples, the seeds of the selected plants with higher Ni content were taken to the growth/germination test in a range of serpentine soils in a growth chamber condition. In order to investigate the rhizosphere microbial diversity, Isatis pinnatiloba and Alyssum dasycarpum which were able to germinate and grow well in the preliminary tests, were introduced to 6-month greenhouse experiment in the range of three serpentine soils with higher bioavailable Ni content. I. pinnatiloba had a better stimulatory effect on the rhizosphere microbial diversity. A total of 22 bacterial isolates were identified from different soil conditions in the end of experiment. Following microbial identification and confirmation tests, 11 isolates were found to be resistant and tolerant to the increasing concentrations of Ni, Pb, Cd and Zn in the range of 50–2,000 mg L − 1, which was considerably higher than those indicated by earlier studies. The strains isolated and identified from the Turkish serpentine soils were the members of genera Arthrobacter, Bacillus, Microbacterium and Staphylococcus.


Phytoremediation Serpentine soils Brassicaceae spp. Nickel Rhizosphere bacteria 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Aboudrar, W., Schwartz, C., Benizri, E., Morel, J. L., & Boularbah, A. (2007). Soil microbial diversity as affected by the rhizosphere of the hyper-accumulator Thlaspi caerulescens under natural conditions. International Journal of Phytoremediation, 9, 41–52.CrossRefGoogle Scholar
  2. Abou-Shanab, R. A., Angle, J. S., Delorme, T. A., Chaney, R. L., Van Berkum, P., Moawad, H., et al. (2003). Rhizobacterial effects on nickel extraction from soil and uptake by Alyssum murale. N. Phytol, 158(1), 219–224.CrossRefGoogle Scholar
  3. Abou-Shanab, R. A. I., Van Berkum, P., & Angle, J. S. (2007). Heavy metal resistance and genotypic analysis of metal resistance genes in Gram-positive and Gram-negative bacteria present in Ni-rich serpentine soil and in the rhizosphere of Alyssum murale. Chemosphere, 68, 360–367.CrossRefGoogle Scholar
  4. Barzanti, R., Ozino, F., Bazzicalupo, M., Gabbrielli, R., Galardi, F., Gonnelli, C., et al. (2007). Isolation and characterization of endophytic bacteria from the nickel hyper-accumulator plant Alyssum bertolonii. Microbial Ecology, 53, 306–316.CrossRefGoogle Scholar
  5. Blaudez, D., Kohler, A., Martin, F., Sanders, D., & Chalot, M. (2003). Poplar metal tolerance protein 1 confers zinc tolerance and is an oligomeric vacuolar zinc transporter with an essential leucine zipper motif. Plant Cell, 15, 2911–2928.CrossRefGoogle Scholar
  6. Bochner, B. R. (1989). Breathprints at the microbial level (pp. 536–539). ASM News.Google Scholar
  7. Bouyoucos, G. J. (1951). A calibration of the hydrometer for making mechanical analysis of soils. Agronomy Journal, 43, 9.CrossRefGoogle Scholar
  8. Çiçek, I., & Türkoğlu, N. (2005). Urban effects on precipitation in Ankara. Atmosfera, 18(3), 172–186.Google Scholar
  9. Davis, P. H. (1965–1985). Flora of Turkey and East Aegean Islands (Vol. 1–9). University Press, Edinburgh.Google Scholar
  10. Giller, K. E., Witter, E., & McGrath, S. P. (1998). Toxicity of heavy metals to microorganisms and microbial processes in agricultural soils: a review. Soil Biology & Biochemistry, 30, 1389–1414.CrossRefGoogle Scholar
  11. Groves, D. J., & Young, F. E. (1975). Epidemiology of antibiotic and heavy metal resistance in bacteria: Resistance patterns in staphylococci isolated from populations not known to be exposed to heavy metals. Antimicrobial Agents and Chemotherapy, 7, 614–621.Google Scholar
  12. Hossner, L. R. (1996). Dissolution for total elemental analysis. In D. L. Sparks et al. (Eds.) Methods of Soil Analysis: Part 3—Chemical Methods (pp. 46–94). Madison: Soil Science Society of America.Google Scholar
  13. Jackson, M. L. (1962). Soil chemical analysis (pp. 214–222). Englewood Cliffs: Prentice-Hall.Google Scholar
  14. Kalra, Y. P., & Maynard, D. G. (1998). Microwave digestion of plant tissue in an open vessel. In Y. P. Kalra (Ed.) Handbook of Reference Methods for Plant Analysis (pp. 63–67). New York: CRC.Google Scholar
  15. Lindsay, W. L., & Norvell, W. A. (1978). Development of a DTPA soil test for zinc, iron, manganese and copper. Soil Science Society of America Journal, 42, 421–428.CrossRefGoogle Scholar
  16. Mengoni, A., Barzanti, R., Gonneli, C., Gabbrielli, R., & Bazzicalupo, M. (2001). Characterization of nickel-resistant bacteria isolated from serpentine soil. Environmental Microbiology, 3, 691–708.CrossRefGoogle Scholar
  17. Mengoni, A., Grassi, E., Barzanti, R., Biondi, E. G., Gonnelli, C., Kim, C. K., et al. (2004). Genetic diversity of bacterial communities of serpentine soil and of rhizosphere of the nickel hyper-accumulator plant Alyssum bertolonii. Microbial Ecology, 48, 209–217.CrossRefGoogle Scholar
  18. Microbial ID. Inc. (MIDI). (1996). Microbial identification system operating manual. Vers. 6. Newark: Microbial ID.Google Scholar
  19. Miller, I., & Berger, T. (1985). Bacteria identification by gas chromatography of whole cell fatty acids. Hewlett-Packard Gas Chromatography Application Note (pp. 228–238). Alto: Hewlett-Packard.Google Scholar
  20. Mohanty, N., Vass, J., & Demeter, S. (1989). Impairment of photosystem 2 activity at the level secondary quinone electron acceptor in chloroplasts treated with cobalt, nickel and zinc ions. Physiologia Plantarum, 76, 389–390.Google Scholar
  21. Motesharezadeh, B., Savaghebi, Gh. R., Alikhani, H. A., & Mir Seyed Hosseini, H. (2008). Effect of Sunflower and Amaranthus Culture and Application of Inoculants on Phytoremediation of the Soils Contaminated with Cadmium, American-Eurasian. Journal of Agricultural Environmental Science, 4(1), 93–103.Google Scholar
  22. Orcan, N. (2003). Alyssum mughlaei (Brassicaceae), a new species from Southwest Anatolia. Nordic Journal of Botany, 23(6), 703–705.CrossRefGoogle Scholar
  23. Pal, A., Choudhuri, P., Dutta, S., Mukherjee, P. K., & Paul, A. K. (2004). Isolation and characterisation of nickel-resistant microflora from serpentine soils of Andaman. World Journal of Microbiology & Biotechnology, 20, 881–886.CrossRefGoogle Scholar
  24. Pal, A., Dutta, S., Mukherjee, P. K., & Paul, A. K. (2005). Occurrence of heavy metal resistance in microflora from serpentine soil of Andaman. Journal of Basic Microbiology, 45, 207–218.CrossRefGoogle Scholar
  25. Proctor, J., & Woodell, S. R. J. (1975). The ecology of serpentine soils. Advances in Ecological Research, 9, 255–365.CrossRefGoogle Scholar
  26. Puschenreiter, M., Stoger, G., Lombi, E., Horak, O., Wenzel, W. W. (2001). Phytoextraction of heavy metal contaminated soils with Thlaspi goesingense and Amaranthus hybridus: rhizosphere manipulation using EDTA and ammonium sulphate. Journal of Plant Nutrition and Soil Science, 164, 615–621.CrossRefGoogle Scholar
  27. Rajkumar, Y., Ma, M., & Freitas, H. (2009). Isolation and characterization of Ni mobilizing PGPB from serpentine soils and their potential in promoting plant growth and Ni accumulation by Brassica spp. Chemosphere, 75, 719–725.CrossRefGoogle Scholar
  28. Reeves, R. D., Kruckeberg, A. R., Adiguzel, N., & Krämer, U. (2001). Studies on the flora of serpentine and other metalliferous areas of western Turkey. South African Journal of Science, 97, 513–517.Google Scholar
  29. Richards, L. A. (1954). Diagnosis and improvement of saline and alkali soils (p. 160). USDA Handbook 60 USA.Google Scholar
  30. Sasser, M. (1990). Technical note 102. Tracking a strain using the Microbial Identification System. Newark: MIS.Google Scholar
  31. Schlegel, H. G., Cosson, J. P., & Baker, A. J. M. (1991). Ni-hyper-accumulating plants provide a niche for Ni-resistant bacteria. Botanica Acta, 104, 18–25.Google Scholar
  32. Schumann, P., Sproer, C., Burghardt, J., Kovacs, G., & Stackebrandt, E. (1999). Reclassification of the species Kocuria erythromyxa (Brooks and Murray 1981) as Kocuria rosea (FIUgge 1886). International Journal of Systematic Bacteriology, 49, 393–396.CrossRefGoogle Scholar
  33. Sheng, X. F., Xia, J. J., Jiang, C. Y., He, L. Y., & Qian, M. (2008). Characterization of heavy metal-resistant endophytic bacteria from rape (Brassica napus) roots and their potential in promoting the growth and lead accumulation of rape. Environmental Pollution, 156, 1164–1170.CrossRefGoogle Scholar
  34. Sheoran, I. S., Singal, H. R., & Singh, R. (1990). Effect of cadmium and nickel on photosynthesis and the enzymes of the photosynthetic carbon reduction cycle in pigeonpea (Cajanus cajan L.). Photosynthesis Research, 23, 345–351.CrossRefGoogle Scholar
  35. Soil Survey Staff (1993). Soil Survey Manual. USDA Handbook No. 18. Washington, DC: US Government Printing Office.Google Scholar
  36. Stoppel, R. D., & Schlegel, H. G. (1995). Nickel-resistant bacteria from anthropogenically nickel-polluted and naturally nickel-percolated ecosystems. Applied and Environmental Microbiology, 61, 2276–2285.Google Scholar

Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  1. 1.Department of Soil Science, Faculty of AgricultureAnkara UniversityAnkaraTurkey
  2. 2.Department of Plant Protection, Faculty of AgricultureAtaturk UniversityErzurumTurkey
  3. 3.Department of Soil Science, Faculty of AgricultureAtaturk UniversityErzurumTurkey

Personalised recommendations