1887

Abstract

Natural plasmid transformation may be a mechanism for the horizontal transfer of non-conjugative plasmids in the marine environment, yet there are few marine model systems available for the study of this process. Using multimers of IncQ/P4 plasmids and a filter transformation assay, we have measured the effects of nutrients, salinity, temperature, as well as the development and maintenance of competence for genetic transformation in the high frequency of transformation (HFT) marine strain WJT-1C. Transformation frequency was proportional to the amount of DNA used from 0·1 to 1·0 μg DNA and was saturated at concentrations greater than 1·0 μg. Competence began in the early-exponential phase and reached a maximum at the onset of stationary phase. Once attained, competence was maintained in both spent and nutrient-free media for at least 10 d. Thus, the establishment and maintenance of competence was unique compared to previously described transformation systems. Temperatures ranging from 4 to 33 °C had no significant effect on the maximal transformation frequency of WJT-1C, but at 37 °C the transformation frequency was reduced. However, temperature did affect the rate of the transformation process. Salinities in the range 12 to 50% had no significant effect on the transformation frequency but transformation frequencies were lower at 6% and 63%. Cells were transformed equally well in nutrient-free media or rich media. The ability of this marine HFT strain to develop competence under a wide spectrum of conditions and to maintain the competent state indicates that natural plasmid transformation could occur in conditions found in tropical and subtropical estuaries.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-139-4-753
1993-04-01
2021-05-07
Loading full text...

Full text loading...

/deliver/fulltext/micro/139/4/mic-139-4-753.html?itemId=/content/journal/micro/10.1099/00221287-139-4-753&mimeType=html&fmt=ahah

References

  1. Ahlquist E. F., Fewson C. A., Podmore J., Rowell V. 1980; Competence for genetic transformation in Acinetobacter calcoaceticus NCIB8250. FEMS Microbiology Letters 7:107–109
    [Google Scholar]
  2. Albano M., Hahn J., Dubnau D. 1987; Expression of competence genes in Bacillus subtilis. Journal of Bacteriology 169:3110–3117
    [Google Scholar]
  3. Avery T., MacLeod C. M., McCarty M. 1944; Studies on the chemical nature of the substance inducing transformation in pneumococcal types. Journal of Experimental Medicine 79:137–159
    [Google Scholar]
  4. Bale M. J., Fry J. C., Day M. J. 1987; Plasmid transfer between strains of Pseudomonas aeruginosa on membrane filters attached to river stones. Journal of General Microbiology 133:3099–3107
    [Google Scholar]
  5. Bale M. J., Fry J. C., Day M. J. 1988; Transfer and occurrence of large mercury resistance plasmids in river epilithon. Applied and Environmental Microbiology 54:972–978
    [Google Scholar]
  6. Bazalyan V. L., Ayzatullin T. A. 1979; Kinetics of enzymatic hydrolysis of DNA in seawater. Oceanology 19:30–33
    [Google Scholar]
  7. Broda P. 1979 Plasmids Oxford: Freeman;
    [Google Scholar]
  8. Carlson C. A., Stewart G. J., Ingraham J. L. 1985; Thymidine salvage in Pseudomonas stutzeri and Pseudomonas aeruginosa provided by heterologous expression of Escherichia coli thymidine kinase gene. Journal of Bacteriology 163:291–295
    [Google Scholar]
  9. Contente S., Dubnau D. 1979; Characterization of plasmid transformation in Bacillus subtilis: kinetic properties and the effect of DNA conformation. Molecular and General Genetics 167:251–258
    [Google Scholar]
  10. Cruze J. A., Singer J. T., Finnerty W. R. 1979; Conditions for quantitative transformation in Acinetobacter calcoaceticus. Current Microbiology 3:129–132
    [Google Scholar]
  11. DeFlaun M. F., Paul J. H. 1986; Hoechst 33258 staining of DNA in agarose gel electrophoresis. Journal of Microbiological Methods 5:265–270
    [Google Scholar]
  12. DeFlaun M. F., Paul J. H. 1989; Detection of exogenous gene sequences in dissolved DNA from aquatic environments. Microbial Ecology 18:21–28
    [Google Scholar]
  13. DeFlaun M. F., Paul J. H., Jeffrey W. H. 1987; Distribution and molecular weight of dissolved DNA in subtropical estuarine and oceanic environments. Marine Ecology Progress Series 38:65–73
    [Google Scholar]
  14. Felsenstein K. M. 1988; A modified protocol for the pZ523 columns: a legitimate alternative for plasmid DNA. BioTechniques 6:847–848
    [Google Scholar]
  15. Foster T. S. 1983; Plasmid-determined resistance to antibiotics and toxic heavy metal ions in bacteria. Microbiological Reviews 47:361–409
    [Google Scholar]
  16. Frey J., Bagdasarian M. 1989; The molecular biology of IncQ plasmids. In Promiscuous Plasmids of Gram-negative Bacteria pp. 79–94, Edited by. Thomas. C. M. London: Academic Press;
    [Google Scholar]
  17. Frischer M. E., Thurmond J. M., Paul J. H. 1990; Natural plasmid transformation in a high-frequency-of-transformation marine Vibrio strain. Applied and Environmental Microbiology 56:3439–3444
    [Google Scholar]
  18. Fry J. C., Day M. J. 1990; Plasmid transfer in the epilithon. In Bacterial Genetics in Natural Environments pp. 55–80, Edited by. Fry J. C., Day M. J. London: Chapman & Hall;
    [Google Scholar]
  19. Fulthorpe R. R., Wyndham R. C. 1991; Transfer and expression of the catabolic plasmid pBRC60 in wild bacterial recipients in a freshwater ecosystem. Applied and Environmental Microbiology 57:1546–1553
    [Google Scholar]
  20. Graham B. J., Istock C. A. 1978; Genetic exchange in Bacillus subtillis in soil. Molecular and General Genetics 166:287–290
    [Google Scholar]
  21. Griffith O. M. 1988; Large-scale isolation of plasmid DNA using high speed centrifugation methods. BioTechniques 6:725–727
    [Google Scholar]
  22. Henschke R. B., Schmidt F. R. J. 1990; Plasmid mobilization from genetically engineered bacteria to members of the indigenous soil microflora in situ. Current Microbiology 20:105–110
    [Google Scholar]
  23. Jeffrey W. H., Paul J. H., Stewart G. J. 1990; Natural transformation of a marine Vibrio species by plasmid DNA. Microbial Ecology 19:259–269
    [Google Scholar]
  24. Juni E. 1972; Interspecies transformation of Acinetobacter: genetic evidence for a ubiquitous genus. Journal of Bacteriology 112:917–931
    [Google Scholar]
  25. Klingmuller W. 1991; Plasmid transfer in natural soil: a case by case study with nitrogen-fixing Enterobacter. FEMS Microbiology Ecology 85:107–116
    [Google Scholar]
  26. Lacks S., Greenberg B. 1973; Competence for deoxyribonucleic acid uptake and deoxyribonuclease action external to cells in the genetic transformation of Diplococcus pneumoniae. Journal of Bacteriology 114:152–163
    [Google Scholar]
  27. Maeda M., Taga N. 1974; Occurrence and distribution of deoxyribonucleic acid hydrolyzing bacteria in seawater. Journal of Experimental Marine Biology and Ecology 14:157–169
    [Google Scholar]
  28. Meyer R., Laux R., Boch G., Hinds M., Bayly R., Shapiro J. A. 1982; Broad-host range IncP-4 plasmid R1162: effects of deletions and insertions on plasmid maintenance and host range. Journal of Bacteriology 152:140–150
    [Google Scholar]
  29. Morita R. Y. 1985; Starvation and miniaturization of heterotrophs, with special emphasis on maintenance of the starved viable state. In Bacteria in Their Natural Environments pp. 111–130, Edited by. Fletcher M. M., Floodgate. G. D. London: Academic Press;
    [Google Scholar]
  30. Norelli J. L., Burr T. J., Adriana A. M., Cicero L., Gilbert M. T., Katz B. H. 1991; Homologous streptomycin resistance gene present among diverse Gram-negative bacteria in New York State apple orchards. Applied and Environmental Microbiology 57:486–491
    [Google Scholar]
  31. Novick R. P. 1969; Extrachromosomal inheritance in bacteria. Bacteriological Reviews 33:210–263
    [Google Scholar]
  32. Novitsky J. A. 1986; Degradation of dead microbial biomass in a marine sediment. Applied and Environmental Microbiology 52:504–508
    [Google Scholar]
  33. Ogram A., Sayler G. S., Barkay T. 1987; The extraction and purification of microbial DNA from sediments. Journal of Microbiological Methods 7:57–66
    [Google Scholar]
  34. Pakula R., Walczak W. 1963; On the nature of competence of transformable streptococci. Journal of General Microbiology 31:125–133
    [Google Scholar]
  35. Paul J. H. 1982; The use of Hoechst dyes 33258 and 33342 for enumeration of attached and planktonic bacteria. Applied and Environmental Microbiology 43:939–944
    [Google Scholar]
  36. Paul J. H., Jeffrey W. H., DeFlaun M. F. 1987; Dynamics of extracellular DNA in the marine environment. Applied and Environmental Microbiology 53:170–179
    [Google Scholar]
  37. Paul J. H., Frischer M. E., Thurmond J. M. 1991; Gene transfer in marine water column and sediment microcosms by natural plasmid transformation. Applied and Environmental Micro-biology 57:1509–1515
    [Google Scholar]
  38. Paul J. H., Thurmond J. M., Frischer M. E., Cannon J. P. 1992; Intergeneric natural plasmid transformation between E. coli and a marine Vibrio species. Molecular Ecology 1:37–46
    [Google Scholar]
  39. Pretorius-Guth I. M., Puhler A., Simon R. 1990; Conjugal transfer of megaplasmid 2 between Rhizobium meliloti strains in alfalfa nodules. Applied and Environmental Microbiology 56:2354–2359
    [Google Scholar]
  40. Redfield R. J. 1991; sxy-1, a Haemophilus influenzae mutation causing greatly enhanced spontaneous competence. Journal of Bacteriology 173:5612–5618
    [Google Scholar]
  41. Rochelle P. A., Day M. J., Fry J. C. 1988; Occurrence, transfer and mobilization in epilithic strains of Acinetobacter of mercury-resistance plasmids capable of transformation. Journal of General Microbiology 134:2933–2941
    [Google Scholar]
  42. Rochelle P. A., Fry J. C., Day M. J. 1989; Factors affecting conjugal transfer of plasmids encoding mercury resistance from pure cultures and mixed natural suspensions of epilithic bacteria. Journal of General Microbiology 135:409–424
    [Google Scholar]
  43. Sambri B., Lovett M. A. 1989; Influence of several experimental parameters on inter- and intra-specific mating process efficiency. Microbiologica 12:323–328
    [Google Scholar]
  44. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning, a Laboratory Manual, 2nd edn.. Cold Spring Harbor, NY:: Cold Spring Harbor Laboratory.;
    [Google Scholar]
  45. Saunders J. R., Saunders V. A. 1988; Bacterial transformation with plasmid DNA. Methods in Microbiology 21:79–128
    [Google Scholar]
  46. Saye D. J., Ogunseitan O. A., Sayler G. S., Miller R. V. 1990; Transduction of linked chromosomal genes between Pseudomonas aeruginosa strains during incubation in situ in a freshwater habitat. Applied and Environmental Microbiology 56:140–145
    [Google Scholar]
  47. Schutt C. 1990; Plasmids and their role in natural aquatic bacterial communities. In Aquatic Microbial Ecology: Biochemical and Molecular Approaches pp. 160–183, Edited by. Overbeck G., Chrost R. J. New York: Springer-Verlag;
    [Google Scholar]
  48. Seifert H. S., Ajioka R. S., Marchal C., Sparling P. F., Magdalene So. 1988; DNA transformation leads to pilin antigenic variation in Neisseria gonorrhoeae. Nature; London: 336392–395
    [Google Scholar]
  49. Smit E., VanElsas J. D., VanVeen J. A., DeVos W. M. 1991; Detection of plasmid transfer from Pseudomonas fluorescens to indigenous bacteria in soil using bacteriophage OR2f for donor counter selection. Applied and Environmental Microbiology 57:3482–3488
    [Google Scholar]
  50. Smith H. O., Danner D. B., Deich R. A. 1981; Genetic transformation. Annual Review of Biochemistry 50:140–145
    [Google Scholar]
  51. Smith J. M., Dowson C. J., Spratt B. G. 1991; Localized sex in bacteria. Nature; London: 34929–31
    [Google Scholar]
  52. Sparling P. F. 1966; Genetic transformation of Neisseria gonorrhoeae to streptomycin resistance. Journal of Bacteriology 92:1364–1371
    [Google Scholar]
  53. Stewart G. J. 1989; The mechanism of natural transformation. In Gene Transfer in the Environment pp. 139–164, Edited by. Levy S. B., Miller R. V. New York: McGraw Hill;
    [Google Scholar]
  54. Stewart G. J., Carlson C. A. 1986; The biology of natural transformation. Annual Review of Microbiology 40:211–235
    [Google Scholar]
  55. Stewart G. J., Cyr D. H. 1987; Distribution of natural transformation ability among marine bacteria (abstract no. 22A-04 1435H). EOS 68:1712
    [Google Scholar]
  56. Stewart G. J., Sinigalliano C. D. 1990; Detection of horizontal gene transfer by natural transformation in native and introduced species of bacteria in marine and synthetic sediments. Applied and Environmental Microbiology 56:1818–1824
    [Google Scholar]
  57. Tomasz A., Hotchkiss R. D. 1964; Regulation of the transformability of pneumococcal cultures by macromolecular cell products. Proceedings of the National Academy of Sciences of the United States of America 51:480–487
    [Google Scholar]
  58. Trevors J. T., Oddie K. M. 1986; R-plasmid transfer in soil and water. Canadian Journal of Microbiology 32:610–613
    [Google Scholar]
  59. VanElsas J. D., Nikkel M., Van Overbeek L. S. 1989; Detection of plasmid RP4 transfer in soil and rhizosphere, and the occurrence of homology to RP4 in soil bacteria. Current Microbiology 19:375–381
    [Google Scholar]
  60. Zar J. H. 1984 Biostatistical Analysis pp. 162–209 New Jersey: Prentice Hall;
    [Google Scholar]
  61. Zervos P. H., Morris L. M., Hell Wig R. J. 1988; A novel method for rapid isolation of plasmid DNA. BioTechniques 6:238–242
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-139-4-753
Loading
/content/journal/micro/10.1099/00221287-139-4-753
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error