Transfer of broad-host-range antibiotic resistance plasmids in soil microcosms

Abstract

Broad-host-range plasmids, belonging to IncP (RP4 and pUPI102) and IncC (R57.b), were studied for intrageneric and intergeneric gene transfer in three different soil microcosms. RP4 was transferred intragenerically in clay loam, sandy loam, and sandy microcosms at frequencies of 0.71×10−2, 0.83×10−2, and 0.41×10−2 respectively, optimally at 37°C and at 100% vol/wt moisture content. Under similar conditions, R57.b was also transferred at frequencies of 0.38×10−2, 0.58×10−2, and 0.80×10−5 respectively at 30°C. Both RP4 and R57.b were transferred at low frequency at 20°C. Kinetics of plasmid transfer revealed that 48 h was the optimum time for intrageneric conjugal gene transfer. Gene transfer frequency was tenfold higher in all nutrient-amended soil microcosms than in the absence of nutrient amendment. RP4 was transferred to an indigenous soil bacteriumBeijerinckia indica in a nonsterile soil microcosm and to other indigenous soil bacteria, viz.Xanthomonas campestris, Azotobacter chroococcum, Acinetobacter calcoaceticus, Achromobacter agili, andRhizobium meliloti in sterile soil microcosms. pUPI102 was transferred fromA. calcoaceticus BD413 toEscherichia coli K12 J53 at a frequency of 0.75×10−6 and 1.1×10−6 in clay loam and sandy loam microcosms respectively. However, no gene transfer was observed in any soil microcosm when strains ofA. calcoaceticus BD413 (pUPI102) andE. coli K12 J53.2 (RP4) were used for conjugal mating. Plasmid RP4 was found to be 100% stable in all the above microorganisms.

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Literature Cited

  1. 1.

    Bachmann BJ (1972) Pedigrees of some mutant strains ofE. coli K12. Bacterial Rev 36:525–527

    Google Scholar 

  2. 2.

    Berg G, Trevors JT (1990) Bacterial conjugation betweenE. coli andPseudomonas spp. donor and recipients cells in soil. J Ind Microbiol 5:79–84

    Google Scholar 

  3. 3.

    Chopade BA (1986) Genetics of antibiotic resistance inAcinetobacter calcoaceticus. Ph.D. Thesis, University of Nottingham, England

    Google Scholar 

  4. 4.

    Chopade BA, Wise PJ, Towner KJ (1985) Plasmid transfer and behaviour inAcinetobacter calcoaceticus EBF 65/65. J Gen Microbiol 131:2805–2811

    Google Scholar 

  5. 5.

    Chopade BA, Patwardhan RB, Dhakephalkar PK (1993)Acinetobacter infections in India: genetic and molecular biological studies and some approaches to the problem. In: Proceedings of tropical diseases: molecular biology and control strategies. Lucknow: Central Drug Research Institute, CSIR publications, New Delhi, in press

    Google Scholar 

  6. 6.

    Datta N, Hedges RW (1972) R factors identified in Paris, some conferring gentamycin resistance, constitute a new compatibility group. Ann Microbiol (Paris) 123:849–859

    Google Scholar 

  7. 7.

    Datta N, Hedges RW, Shaw EJ, Sykes RB, Richmond MH (1971) Properties of R factors fromPseudomonas aeruginosa. J Bacteriol 108:1244–1249

    Google Scholar 

  8. 8.

    Deshpande LM, Kapadnis BP, Chopade BA (1993) Metal resistance inAcinetobacter and its relation to β-lactamase production. BioMetals 6:55–59

    Google Scholar 

  9. 9.

    Dhakephalkar PK, Murthi S, Seshadri PG, Puntambekar M, Chopade BA (1992) Genetic studies on pUPI102, a plasmid encoding resistance to tetracycline, gentamycin, neomycin and mercury inA. juni ACN4 In: Proceedings of DAE symposium on molecular biology of microorganisms. Bombay: Dept. of Atomic Energy, Govt. of India, pp 72–77

    Google Scholar 

  10. 10.

    Juni E (1972) Interspecies transformation ofAcinetobacter: genetic evidence for a ubiquitous genus. J Bacteriol 112:917–931

    Google Scholar 

  11. 11.

    Kanwar JS (1976) Soil fertility theory and practice. New Delhi: Indian Council of Agricultural Research [ICAR]

    Google Scholar 

  12. 12.

    Karasovsky VN, Stotzky G (1987) Conjugation and genetic recombination inE. coli in sterile and non sterile soil. Soil Biol Biochem. 19:631–638

    Google Scholar 

  13. 13.

    Kinkle BK, Schmidt EL (1991) Transfer of pea symbiotic plasmid pJB5JI in non sterile soil. Appl Environ Microbiol 57:3264–3269

    Google Scholar 

  14. 14.

    Narayana N, Shah C (1966) Physical properties of soils. Bombay: Manaktalas Publishers

    Google Scholar 

  15. 15.

    Prosser JI, Reid N (1992) A mathematical model of gene transfer on surfaces. Abstracts of 6th International Symposium on Microbial Ecology, Barcelona, p 105

  16. 16.

    Ramos MI, Gonzalez JM (1991) Conjugation transfer of recombinant DNA in cultures and in soils. Appl Environ Microbiol 57:3020–3027

    Google Scholar 

  17. 17.

    Schilf W, Klingmuller W (1983) Experiments withE. coli on the dispersal of plasmids in environmental samples. Recomb DNA Tech Bull 6:101–102

    Google Scholar 

  18. 18.

    Scott EM, Van Elsas JD, Bazin MJ, Lynch JM (1992) A mathematical model of gene exchange in a soil environment. Abstracts of 6th International Symposium on Microbial Ecology, Barcelona, p 105

  19. 19.

    Silhavy TJ, Berman ML, Erquist LW (1984) In: Experiments with gene fusions. Cold Spring Harbor Laboratory, NY: Cold Spring Harbor Laboratory Press, pp 147–148

    Google Scholar 

  20. 20.

    Smith MS, Thomas GW, White RE, Ritonga D (1985) Transport ofE. coli through intact and disturbed soil columns. J Environ Qual 14:87–91

    Google Scholar 

  21. 21.

    Stotzky G, Babich H (1986) Survival of and genetic transfer by genetically engineered bacteria in natural environments. Adv Appl Microbiol 12:547–563

    Google Scholar 

  22. 22.

    Stotzky G, Rem LT (1966) Influence of clay minerals on microorganisms. Can J Microbiol 12:547–633

    Google Scholar 

  23. 23.

    Sun L, Bazin MJ, Lynch JM (1993) Plasmid dynamics in a model soil column. Mol Ecol 2:9–15

    Google Scholar 

  24. 24.

    Taher Khalil A, Gealt MA (1987) Temperature, pH and cations affect the ability ofE. coli to mobilize plasmids in broth and synthetic waste water. Can J Microbiol 33:733–737

    Google Scholar 

  25. 25.

    Trevors JT, Barkay T (1987) Gene transfer among bacteria in soil and acquatic environments. Can J Microbiol 33:191–196

    Google Scholar 

  26. 26.

    Trevors JT, Berg G (1988) Conjugal RP4 transfer betweenPseudomonas in soil and recovery of RP4 plasmid DNA from soil. Syst Appl Microbiol 11:223–227

    Google Scholar 

  27. 27.

    Trevors JT, Oddie KM (1986) R plasmid transfer in soil and water. Can J Microbiol 32:610–613

    Google Scholar 

  28. 28.

    Van Elsas JD, Johana M (1987) Transfer of plasmid pFT30 between bacilli in soil as influenced by bacterial population dynamics and soil conditions. Soil Biol Biochem 19:639–647

    Google Scholar 

  29. 29.

    Weinberg SR, Stotzky G (1972) Conjugation and genetic recombination ofE. coli in soil. Soil Biol Biochem 4:171–180

    Google Scholar 

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Correspondence to Dr. B. A. Chopade.

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Naik, G.A., Bhat, L.N., Chopade, B.A. et al. Transfer of broad-host-range antibiotic resistance plasmids in soil microcosms. Current Microbiology 28, 209–215 (1994). https://doi.org/10.1007/BF01575963

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Keywords

  • Acinetobacter
  • Clay Loam
  • Azotobacter
  • Soil Microcosm
  • Nonsterile Soil