1887

Abstract

This study investigated mobilization and persistence of the recombinant, catabolic plasmid pD10 in laboratory microcosms by natural mobilizing plasmids, recently isolated from epilithic bacteria by exogenous isolation in , as opposed to their isolation in the strains in which they occur . Initial experiments in simple, beaker microcosms were used to optimize conditions for selection of the donor, recipient and transconjugant populations. Studies in a recirculating stream microcosm showed that donor, KT2440(pD10, pQKH6), and recipient, UWC6, populations, although declining with time, were able to survive for three weeks and that colonization and survival was predominantly in the epilithon of the microcosm rather than in the liquid phase. Transconjugants UWC6(pD10, pQKH6) were also isolated from epilithon, showing that plasmid pD10 had been mobilized by plasmid pQKH6. The donor strain was able to survive at 100-fold greater numbers than the recipient, but transconjugants were not isolated after day 15. Separate inoculation of the donor and recipient strains into the microcosm showed that they were able to colonize other stones, where transconjugants then arose as a direct result of plasmid transfer within the epilithon. A catechol 2,3-dioxygenase gene, , was used to facilitate identification of donors and transconjugants. Preliminary laboratory matings showed that pD10 transferred to indigenous epilithic strains identified as and . These results suggest that in the absence of selection, mobilization of introduced recombinant genes encoded by pD10 occurs at easily detectable frequencies, even in an oligotrophic environment.

Loading

Article metrics loading...

/content/journal/micro/10.1099/13500872-140-7-1555
1994-07-01
2024-04-24
Loading full text...

Full text loading...

/deliver/fulltext/micro/140/7/mic-140-7-1555.html?itemId=/content/journal/micro/10.1099/13500872-140-7-1555&mimeType=html&fmt=ahah

References

  1. Awong J., Bitton G., Chaudry G.R. Microcosm for assessing survival of genetically engineered microorganisms in aquatic environments. Appl Environ Microbiol 1990; 56:977–983
    [Google Scholar]
  2. Bagdasarian M., Lurz R., Riickert B., Franklin F.C.H., Bagdasarian M.M., Frey J., Timmis K.N. Specific purpose plasmid cloning vectors II Broad-host range, high copy number, RSFIOIO-derived vectors, and a host-vector system for gene cloning in Pseudomonas. Gene 1981; 16:237–247
    [Google Scholar]
  3. Bale M.J., Fry J.C., Day M.J. Transfer and occurrence of large mercury resistance plasmids in river epilithon. Appl Environ Microbiol 1988a; 54:972–978
    [Google Scholar]
  4. Bale M.J., Fry J.C., Day M.J. Novel method for studying plasmid transfer in undisturbed river epilithon. Appl Environ Microbiol 1988b; 54:2756–2758
    [Google Scholar]
  5. Bartels I., Knackmuss H.J., Reineke W. Suicide inactivation of catechol 2,3-dioxygenase from Pseudomonas putida mt-2 by 3-halocatechols. Appl Environ Microbiol 1984; 47:500–505
    [Google Scholar]
  6. Bradley D.E. Determination of pili by conjugative bacterial drug resistance plasmids of incompatibility groups B, C, H, J, K, M, V and X. J Bacteriol 1980; l41:828–837
    [Google Scholar]
  7. Bradley D.E. Specification of the conjugative pili and surface mating systems of Pseudomonas plasmids. J Gen Microbiol 1983; 129:2545–2556
    [Google Scholar]
  8. Bradley D.J. Gene transfer in the epilithon of river stones 1992 PhD thesis, University of Wales, Cardiff, UK;
    [Google Scholar]
  9. Chaudry G.R., Chapalamadugu S. Biodegradation of halogenated organic compounds. Microbiol Rev 1991; 55:59–79
    [Google Scholar]
  10. Contreras A., Molin S., Ramos J.L. Conditional-suicide containment system for bacteria which mineralize aromatics. Appl Environ Microbiol 1991; 57:1504–1508
    [Google Scholar]
  11. Day M.J., Fry J.C., Diaper I. Effect of host cell physiology on plasmid transfer in river epilithon. In Gene Transfers and Environment 1992 Edited by Gauthier M.J. Berlin: pp 35–50
    [Google Scholar]
  12. Fry J., G, Day M.J., Cousland B. Transfer, survival and spread of genetically manipulated organisms (GMOs) in river sediments, soil and agricultural environments. In Biotechnology R & D in the EC vol. II, Detailed Final Report of BAP Contractors 1991 Edited by Vassarotti A., Magnien E. Amsterdam: Elsevier;
    [Google Scholar]
  13. Geesey G.G., Richardson W.T., Yeoman H.G., Irvin R.T., Costerton J.W. Microscopic examination of natural sessile bacterial populations from an alpine stream. Can J Microbiol 1977; 23:1733–1736
    [Google Scholar]
  14. Genthner F., Chatterjee J.P., Barkay T., Bourquin A.W. Capacity of aquatic bacteria to act as recipients of plasmid DNA. Appf Environ Microbiol 1988; 54:115–117
    [Google Scholar]
  15. Gerchakov S.M., Hatcher P.G. Improved technique for analysis of carbohydrates in sediments. Eimnol Oceanog 1972; 17:938–943
    [Google Scholar]
  16. Henschke R.B., Schmidt F.R.I. Plasmid mobilization from a genetically engineered bacteria to members of the indigenous soil microflora in situ. Curr Microbial 1990; 20:105–110
    [Google Scholar]
  17. Hill K.E., Weightman A.J., Fry J.C. Isolation and screening of plasmids from epilithic bacteria which mobilize recombinant plasmid pDlO. Appl Environ Microbiol 1992; 58:1292–1300
    [Google Scholar]
  18. Kado C.I., Liu S.T. Rapid procedure for detection and isolation of large and small plasmids. J Bacteriol 1981; 145:1365–1373
    [Google Scholar]
  19. Klein T.M., Alexander M. Bacterial inhibitors in lake water. Appl Environ Microbiol 1986; 52:114–118
    [Google Scholar]
  20. Liang L.N., Sinclair J.L., Mallory L., Alexander M. Fate in model ecosystems of microbial species of potential use in genetic engineering. Appl Environ Microbiol 1982; 44:708–714
    [Google Scholar]
  21. McClure N.C., Weightman A.J., Fry J.C. Survival of Pseudomonas putida UWC1 containing cloned catabolic genes in a model activated-sludge system. Appl Environ Microbiol 1989; 55:2627–2634
    [Google Scholar]
  22. McClure N.C., Fry J.C., Weightman A.J. Gene transfer in activated sludge. In Bacterial Genetics in Natural Environments 1990a Edited by Fry J.C., Day M.J. London: Chapman & Hall; pp 111–129
    [Google Scholar]
  23. McClure N.C., Saint C.P., Fry J.C., Weightman A.J. The construction of broad-host range genetic markers and their use in monitoring the release of catabolic GEMs to aquatic environments. In 5th European Congress on Biotechnology Proceedings 1990b Edited by Christiansen C., Munck L., Villadsen J. Copenhagen: Munksgaard; 1 pp 169–172
    [Google Scholar]
  24. McClure N.C., Venables W.A. pTDNl, a catabolic plasmid involved in aromatic amine catabolism in Pseudomonas putida mt-2. J Gen Microbiol 1987; 133:2073–2077
    [Google Scholar]
  25. McPherson P., Gealt M.A. Isolation of indigenous wastewater bacterial strains capable of mobilizing plasmid pBR325. Appl Environ Microbiof 1986; 51:904–909
    [Google Scholar]
  26. Morgan J.A.W., Rhodes G., Pickup R.W. Survival of non-culturable Aeromonas salmonicida in lake water. Appl Environ Appl Environ Microbiol 1993; 51:904–909
    [Google Scholar]
  27. Perkins E.J., Gordon M.P., Caceres O., Lurquin P.F. Organisation and sequence analysis of the 2,4-dichlorophenol hydrolase and dichlorocatechol oxidative operons of plasmid pJP4. J Bacteriol 1990; 172:2351–2359
    [Google Scholar]
  28. Ramos J.L., Duque E., Ramos-Gonzalez M. Survival in soils of an herbicide-resistant Pseudomonas putida strain bearing a recombinant TOL plasmid. Appl Environ Microbiol 1991; 57:260–266
    [Google Scholar]
  29. Reasonner D.J., Geldriech E.E. A new medium for the enumeration and subculture of bacteria from potable water. Appl Environ Microbiol 1985; 49:1–7
    [Google Scholar]
  30. Reineke W., Knackmuss H.J. Hybrid pathway for chlorobenzoate metabolism in Pseudomonas sp B13 derivatives. J Bacteriol 1980; 142:467–473
    [Google Scholar]
  31. Rochelle P.A., Fry J.C., Day M.J. An accurate method for estimating sizes of small and large plasmids and DN A fragments by gel electrophoresis. J Gen Microbiol 1986; 132:53–59
    [Google Scholar]
  32. Sandt C.H., Herson D.S. Mobilization of the genetically engineered plasmid pHSV106 from Escherichia coli HB101(pHSV106) to Enterobacter cloacae in drinking water. Appl Environ Microbiol 1991; 57:194–200
    [Google Scholar]
  33. Scheuerman P.R., Schmidt J.P., Alexander M. Factors affecting the survival and growth of bacteria introduced into lake water. Arch Microbiol 1988; 150:320–325
    [Google Scholar]
  34. Sinclair J.L., Alexander M. Role of resistance to starvation in bacterial survival in sewage and lake water. Appl Environ Microbiol 1984; 48:410–415
    [Google Scholar]
  35. Slater J.H., Lovatt D., Weightman A.J., Senior E., Bull A.T. The growth of Pseudomonas putida on chlorinated aliphatic acids and its dehalogenase activity. J Gen Microbiol 1979; 114:125–136
    [Google Scholar]
  36. Smit E., Van Elsas J.D., Van Veen J.A., De Vos W.M. Detection of plasmid transfer from Pseudomonas fluorescens to indigenous bacteria in soil by using bacteriophage R2f for donor counterselection. Appl Environ Microbiol 1991; 57:3482–3488
    [Google Scholar]
  37. Smit E., Venne D., Van Elsas J.D. Mobilization of a recombinant IncQ plasmid between bacteria on agar and in soil via cotransfer or retrotransfer. Appl Environ Microbiol 1993; 59:2257–2263
    [Google Scholar]
  38. Sugino A., Peebles C.L., Kreuzer K.N., Cozzarelli N.R. Mechanism of action of nalidixic acid: purification of Escherichia coli nalA gene product and its relationship to DNA gyrase and a novel nicking-closing enzyme. Proc Natl Acad Sci USA 1977; 74:4767–4771
    [Google Scholar]
  39. Vogel S., Labarbera M. Simple flow tanks for research and teaching. Bioscience 1978; 28:638–643
    [Google Scholar]
  40. Weightman A.J., Don R.H., Lehrbach P.R., Timmis K.N. The identification and cloning of genes encoding haloaromatic catabolic enzymes and the construction of hybrid pathways for substrate mineralization. In Genetic Control of En-vironmental Pollutants 1984 Edited by Omenn G.S., Hollaender A. New York and London: Plenum Press; pp 47–80
    [Google Scholar]
  41. Winstanley C., Morgan J.A.W., Pickup R.W., Jones J.G., Saunders J.R. Differential regulation of lambda pL and pR promoters by a cI repressor in a broad-host range thermoregulated plasmid marker system. Appl Environ Microbiol 1989; 55:771–777
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/13500872-140-7-1555
Loading
/content/journal/micro/10.1099/13500872-140-7-1555
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