Methods for the Detection, Identification, and Enumeration of Microbes

  • David J. Drahos
Part of the Brock/Springer Series in Contemporary Bioscience book series (BROCK/SPRINGER)


The greatest challenge a researcher usually faces when designing definitive experiments to measure the dynamics of microbial populations in the phyllosphere is to develop reliable and practical means to detect, identify, or enumerate a particular strain or group of interest in an environmental sample. While a number of sensitive, and often quite sophisticated and elegant, tools now exist for the task, each has advantages and disadvantages which must be fully appreciated and considered prior to settling on a particular strategy. Failure to take this step early can readily result in erroneous conclusions or an unexpected drain on resources and time. This chapter provides an overview of the current methodologies which have been or could be applied to phyllosphere microbiology, and it includes sufficient key references to allow a more comprehensive assessment of applicability to a particular research plan.


Nalidixic Acid Polymerase Chain Reaction Method Environmental Microbiology Xanthomonas Campestris Nucleic Acid Hybridization 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Adams, M.R., Grubb, S.M., Hamer, A., and Clifford, M.N. 1990. Colorimetrie enumeration of Escherichia coli based on β-glucuronidase activity. Applied and Environmental Microbiology 56:2021–2024.PubMedGoogle Scholar
  2. Andrews, J.H. 1986. How to track a microbe, pp. 14–34 in Fokkema, N.J. and van den Henvel, J. (editors), Microbiology of the Phyllosphere. Cambridge University Press, Cambridge.Google Scholar
  3. Atlas, R.M. 1982. Enumeration and estimation of biomass of microbial components in the biosphere, pp. 84–102 in Burns, R.G., and Slater, J.H. (editors), Experimental Microbial Ecology. Blackwell Scientific Publishers, Oxford.Google Scholar
  4. Atlas, R.M. and Sayler, G.S. 1988. Tracking microorganisms and genes in the environment, pp. 31–45 in Omenn, G.S. (editor), Environmental Biotechnology: Reducing Risks from Environmental Chemicals through Biotechnology. Plenum Press, New York.Google Scholar
  5. Atlas, R.M. and Steffan, R.J. 1988. Target DNA amplification, pp. 224–226 in Sussman, M., Collins, C.H., Skinner, F.A., and Stewart-Tull, D.E. (editors), The Release of Genetically-engineered Micro-organisms. Academic Press, London.Google Scholar
  6. Barry, G. F. 1986. Permanent insertion of foreign genes into the chromosomes of soil bacteria. Bio/Technology 4:446–449.CrossRefGoogle Scholar
  7. Barry, G. F. 1988a. A broad-host-range shuttle system for gene insertion into the chromosome of Gram-negative bacteria. Gene 71:75–84.PubMedCrossRefGoogle Scholar
  8. Barry, G.F. 1988b. Construction of reporter strains, pp. 211–219 in Sussman, M., Collins, C.H., Skinner F.A., and Stewart-Tull, D.E. (editors), The Release of Genetically-engineered Micro-organisms. Academic Press, London.Google Scholar
  9. Bej, A.K., Steffan, R.J., DiCesare, J., Haff, L., and Atlas, R.M. 1990. Detection of coliform bacteria in water by polymerase chain reaction. Applied and Environmental Microbiology 56:307–314.PubMedGoogle Scholar
  10. Bochner, B.R. and Savageau, M. A. 1977. Generalized indicator plate for genetic, metabolic, and taxonomic studies with microorganisms. Applied and Environmental Microbiology 33:434–444.PubMedGoogle Scholar
  11. Bohlool, B.B. and Schmidt, E.L. 1980. The immunofluorescence approach in microbial ecology. Advances in Microbial Ecology 4:203–241.Google Scholar
  12. Borsheim, K.Y., Bratbak, G., and Heldal, M. 1990. Enumeration and biomass estimation of planktonic bacteria and viruses by transmission electron microscopy. Applied and Environmental Microbiology 56:352–356.PubMedGoogle Scholar
  13. Botstein, D., White, R., Skolnick, M., and Davis, R.W. 1980. Genetic mapping using restriction fragment length polymorphisms. American Journal of Human Genetics 32:314–331.PubMedGoogle Scholar
  14. Brayton, P.R. and Colwell, R.R. 1987. Fluorescent antibody staining method for enumeration of viable environmental Vibrio cholerae O1. Journal of Microbiological Methods 6:309–314.CrossRefGoogle Scholar
  15. Brayton, P.R., Tamplin, M.L., Huq, A., and Colwell, R.R. 1987. Enumeration of Vibrio cholerae O1 in Bangladesh waters by fluorescent-antibody direct viable count. Applied and Environmental Microbiology 53:2862–2865.PubMedGoogle Scholar
  16. Bromfield, E.S.P., Sinha, I.B., and Wolynetz, M.S. 1986. Influence of location, host cultivar, and inoculation on the composition of naturalized populations of Rhizobium meliloti in Medicago sativa nodules. Applied and Environmental Microbiology 51:1077–1084.PubMedGoogle Scholar
  17. Brunel, B., Cleyet-Marel, J.-C., Normand, P., and Bardin, R. 1988. Stability of Bradyrhizobium japonicum inoculants after introduction into soil. Applied and Environmental Microbiology 54:2636–2642.PubMedGoogle Scholar
  18. Chaicumpa, W., Thin-Intra, W., Khusmith, S., Tapchaisri, P., Echeverria, P., Kalambaheti, T., and Chongsa-Nguan, M. 1988. Detection with monoclonal antibody of Salmonella typhi antigen 9 in specimens from patients. Journal of Clinical Microbiology 26:1824–1830.PubMedGoogle Scholar
  19. Chaudhry, G.R., Topranzos, G.A., and Bhatti, A.R. 1989. Novel method for monitoring genetically engineered microorganisms in the environment. Applied and Environmental Microbiology 55:1301–1304.PubMedGoogle Scholar
  20. Chopra, I., Shales, S.W., Ward, M.W., and Wallace, L.J. 1981. Reduced expression of Tn10-mediated tetracycline resistance in Escherichia coli containing more than one copy of the transposon. Journal of General Microbiology 126:45–54.PubMedGoogle Scholar
  21. Cole, M.A. and Elkan, G.H. 1979. Multiple antibiotic resistance in Rhizobium japonicum. Applied and Environmental Microbiology 37:867–870.PubMedGoogle Scholar
  22. Colwell R.R., Brayton, P.R., Grimes, D.J., Roszak, D.B., Huq, S.A., and Palmer, L.M. 1985. Viable but non-culturable Vibrio cholerae and related pathogens in the environment: Implications for release of genetically engineered organisms. Bio/Technology 3:817–820.CrossRefGoogle Scholar
  23. Colwell, R.R., Somerville, C., Knight, I., and Straube, W.L. 1988. Detection and monitoring of genetically engineered micro-organisms, pp. 47–60 in Sussman, M., Collins, C.H., Skinner F.A., and Stewart-Tull, D.E. (editors), The Release of Genetically-engineered Micro-organisms. Academic Press, London.Google Scholar
  24. Cook R.J. and Baker, K.F. 1983. The Nature and Practice of Biological Control of Plant Pathogens. American Phytopathological Society, St. Paul, MN.Google Scholar
  25. Cook, R.J., Weller, D.M., Drahos, D.J., Kovacevich, P.A., Hemming, B.C., Barnes, G., and Peirson, E.L. 1991. Establishment, monitoring, and termination of field tests with genetically altered bacteria applied to wheat for biological control of take-all. In MacKenzie, D.R. (editor), Biotechnology Field Test Results. Academic Press, New York. In press.Google Scholar
  26. Demezas, D.H. and Bottomley, P.J. 1986. Autecology in rhizospheres and nodulating behavior of indigenous Rhizobium trifolii. Applied and Environmental Microbiology 52:1014–1019.PubMedGoogle Scholar
  27. Desmonts, C., Minet, J., Colwell, R., and Cormier, M. 1990. Fluorescent-antibody method useful for detecting viable but nonculturable Salmonella spp. in chlorinated wastewater. Applied and Environmental Microbiology 56:1448–1452.PubMedGoogle Scholar
  28. Diatloff, A. 1977. Ecological studies of root-nodule bacteria introduced into field environments. 6. Antigenic and symbiotic stability in Lotononis rhizobia over a 12-year period. Soil Biology and Biochemistry 9:85–88.CrossRefGoogle Scholar
  29. Drahos, D., Brackin, J., and Barry, G. 1985. Bacterial strain identification by comparative analysis of chromosomal DNA restriction patterns. Phytopathology 75:1381 (Abstract).Google Scholar
  30. Drahos, D.J., Hemming, B.C., and McPherson, S. 1986. Tracking recombinant organisms in the environment: β-galactosidase as a selectable, non-antibiotic marker for fluorescent pseudomonads. Bio/Technology 4:439–443.CrossRefGoogle Scholar
  31. Drahos, D.J., Barry, G.F., Hemming, B.C., Brandt, E.J., Skipper, H.D., Kline, E.L., Kluepfel, D.A., Hughes, T.A., and Gooden, D.T. 1988. Pre-release testing procedures: US field test of a lacZY-engineered soil bacterium, pp. 181–191 in Sussman, M., Collins, C.H., Skinner F.A., and Stewart-Tull D.E. (editors), The Release of Genetically-engineered Micro-organisms. Academic Press, London.Google Scholar
  32. Drahos, D.J., Barry, G.F., Hemming, B.C., Brandt, E. J., Kline, E.L., Skipper, H.D., and Kluepfel, D.A. 1991. Spread and survival of genetically marked bacteria in soil. In Day, J.J. and Fry, J.C. (editors), Environmental Release of Genetically Engineered and Other Microorganisms. Academic Press, London. In press.Google Scholar
  33. Ellis, W.R., Ham, G.E., and Schmidt, E.L. 1984. Persistence and recovery of Rhizobium japonicum inoculum in a field soil. Agronomy Journal 76:573–576.CrossRefGoogle Scholar
  34. Festl, H., Lugwig, W., and Schleifer, K.H. 1986. DNA hybridization probe for the Pseudomonas fluorescens group. Applied and Environmental Microbiology 52:1190–1194.PubMedGoogle Scholar
  35. Giovannoni, S.J., Delong, E.F., Olsen, G.J., and Pace, N.R. 1988. Phylogenetic group-specific oligodeoxynucleotide probes for identification of single microbial cells. Journal of Bacteriology 170:720–726.PubMedGoogle Scholar
  36. Goodnow, R.A., Harrison, M.D., Morris, J.D., Sweeting, K.B., and Laduca, R.J. 1990. Fate of ice nucleation-active Pseudomonas syringae strains in alpine soils and waters and in synthetic snow samples. Applied and Environmental Microbiology 56:2223–2227.PubMedGoogle Scholar
  37. Goodwin, D. and Slater, J.H. 1979. The influence of the growth environment on the stability of a drug resistance plasmid in Escherichia coli K-12. Journal General Microbiology 111:201–210.Google Scholar
  38. Gross, D.C., Cody, Y.S., Proebsting, E.L., Jr., Radamaker, G., and Spotts, R.A. 1984. Ecotypes and pathogenicity of ice nucleation active Pseudomonas syringae isolated from deciduous fruit tree orchards. Phytopathology 74:243–269.CrossRefGoogle Scholar
  39. Hartung, J.S. and Civerolo, E.L. 1987. Genomic fingerprints of Xanthomonas campestris pv. citri strains from Asia, South America, and Florida. Phytopathology 77:282–285.CrossRefGoogle Scholar
  40. Hemming, B.C. and Drahos, D.J. 1984. β-Galactosidase, a selectable non-antibiotic marker for fluorescent pseudomonads. Journal of Cellular Biochemistry, Supplement 8b:252.Google Scholar
  41. Hirano, S.S. and Upper, C.D. 1986. Bacterial nucleation of ice in plant leaves, pp. 730–738 in Parker, I. (editor), Methods in Enzymology. Protons and Water: Structure and Translocation, vol. 127, Academic Press, New York.CrossRefGoogle Scholar
  42. Hirano, S.S., Baker, L.S., and Upper, C.D. 1985. Ice nucleation temperature of individual leaves in relation to population sizes of ice nucleation active bacteria and frost injury. Plant Physiology 77:259–265.PubMedCrossRefGoogle Scholar
  43. Hirano, S.S., Rouse, D.I., Amy, D.C., Nordheim, E.V., and Upper, CD. 1981. Epiphytic ice nucleation active (INA) bacterial populations in relation to halo blight incidence in oats. Phytopathology 71:881.Google Scholar
  44. Hirano, S.S., Rouse, D.I., and Upper, C.D. 1987. Bacteial ice nucleation as a predictor of bacterial brown spot disease on snap beans. Phytopathology 77:1078–1084.CrossRefGoogle Scholar
  45. Holben, W.E., Jansson, J.K., Chelm, G.K., and Tiedje, J.M. 1988. DNA probe method for the detection of specific microorganisms in the soil bacterial community. Applied and Environmental Microbiology 54:703–711.PubMedGoogle Scholar
  46. Holt, J.G. (editor). 1989. Bergey’s Manual of Systematic Bacteriology. Williams & Wilkins, Baltimore, MD. 2648 pp.Google Scholar
  47. Ingram, C., Brawner, M., Youngman, P., and Westpheling, J. 1989. xylE functions as an efficient reporter gene in Streptomyces spp.: Use for the study of galP1, a catabolite-controlled promoter. Journal of Bacteriology 171:6617–6624.PubMedGoogle Scholar
  48. Jefferson, R.A., Burgess, S.M., and Hirsch, D. 1986. β-Glucuronidase from Escherichia coli as a gene-fusion marker. Proceedings of the National Academy of Sciences USA 83:8447–8451.CrossRefGoogle Scholar
  49. Jefferson, R.A., Kavanagh, T.A., and Bevan, M.W. 1987. GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO Journal 6:3901–3907.PubMedGoogle Scholar
  50. Kelch, W.J. and Lee, J.S. 1978. Antibiotic resistance patterns of gram-negative bacteria isolated from environmental sources. Applied and Environmental Microbiology 36:450–456.PubMedGoogle Scholar
  51. Kloepper, J.W. and Schroth, M.N. 1981. Relationship in in vitro antibiosis of plant growth-promoting rhizobacteria to plant growth and the displacement of root microflora. Phytopathology 71:1020–1024.CrossRefGoogle Scholar
  52. Kloepper, J.W., Leong, L., Teintze, M., and Schroth, M.N. 1980. Pseudomonas siderophores: A mechanism explaining disease-suppressive soils. Current Microbiology 4:317–320.CrossRefGoogle Scholar
  53. Kirchner, G., Roberts, J.L., Gustafson, G.D., and Ingolia, T.D. 1989. Active bacterial luciferase from a fused gene: expression of a Vibrio Harveyi lux AB translational fusion in bacteria, yeast and plant cells. Gene 81:349–354.PubMedCrossRefGoogle Scholar
  54. Kogure, K., Simidu, U., and Taga, N. 1979. A tentative direct microscopic method for counting living marine bacterai. Canadian Journal of Microbiology 25:415–420.PubMedCrossRefGoogle Scholar
  55. Kogure, K., Simidu, U., Taga, N., and Colwell, R.R. 1987. Correlation of direct viable counts with heterotrophic activity for marine bacteria. Applied and Environmental Microbiology 53:2332–2337.PubMedGoogle Scholar
  56. Koncz, C., Olsson, O., Langridge, W.R., Schell, J., and Szalay, A. 1987. Expression and assembly of functional bacterial luciferase in plants. Proceedings of the National Academy of Sciences USA 84:131–135.CrossRefGoogle Scholar
  57. Leary, J.J., Brigati, D.J., and Ward, D.C. 1983. Rapid and sensitive colorimetric method for visualizing biotin-labeled DNA probes hybridized to DNA or RNA immobilized on nitrocellulose: Bio-blots. Proceedings of the National Academy of Sciences USA 80:4045–4049.CrossRefGoogle Scholar
  58. Legocki, R.P., Legocki, M, Baldwin, T.O., and Szalay, A.A. 1987. Bioluminescence in root nodules of soybean controlled by nitrogenase nifD and nifH promoters, pp. 282–287 in Verma, D.P.S. and Bisson, N. (editors), Molecular Genetics of Plant-Microbe Interactions. Martinus Nijhoff Publications, Dordrecht, The Netherlands.Google Scholar
  59. Levy, S.B. 1984. Resistance to the tetracyclines, pp. 191–240 in Bryan, L.E. (editor), Antimicrobial Drug Resistance. Academic Press, Orlando, FL.Google Scholar
  60. Lindgren, P.B., Frederick, R., Govindarajan, A.G., Panopoulos, N.J., Staskawicz, B.J., and Lindow, S.E. 1989. An ice nucleation reporter gene system: identification of inducible pathogenicity genes in Pseudomonas syringae pv. phaseolicola. The EMBO Journal 8:1291–1301.PubMedGoogle Scholar
  61. Lindow, S.E., Arny, D.C., Barchet, W.R., and Upper, C.D. 1978a. The role of bacterial ice nuclei in frost injury to sensitive plants, pp. 249–261 in Li, P.H. and Sakai, A. (editors), Plant Cold Hardiness and Freezing Stress. Academic Press, New York.Google Scholar
  62. Lindow, S.E., Amy, D.C., and Upper, C.D. 1978b. Erwinia herbicola: an active ice nucleus incites frost damage to maize. Phytopathology 68:523–527.CrossRefGoogle Scholar
  63. Lindow, S.E., Amy, D.C., and Upper, C.D. 1978c. Distribution of ice nucleation active bacteria on plants in nature. Applied and Environmental Microbiology 36:831–838.PubMedGoogle Scholar
  64. Lindstrom, K., Lipsanen, P., and Kaifalainen, S. 1990. Stability of markers used for identification of two Rhizobium galegae inoculant strains after five years in the field. Applied and Environmental Microbiology 56:444–450.PubMedGoogle Scholar
  65. Lipsanen, P. and Lindstrom, K. 1989. Lipopolysaccharide and protein profiles of Rhizobium sp. (Galega) strains. FEMS Microbiological Letters 58:323–328.CrossRefGoogle Scholar
  66. Macario, A.J.L. and Conway de Macario, E. (editors). 1985. Monoclonal Antibodies Against Bacteria. Academic Press, Orlando, FL.Google Scholar
  67. Maki, R.L., Galyan, E.L., Chang-Chien, M., and Caldwell, D.R. 1974. Ice nucleation induced by Pseudomonas syringae. Applied and Environmental Microbiology 28:456–460.Google Scholar
  68. Makkar, N.S. and Casida, L.E. 1987. Technique for estimating low numbers of a bacterial strain(s) in soil. Applied and Environmental Microbiology 53:887–888.PubMedGoogle Scholar
  69. Malvick, D.K. and Moore, L.W. 1988. Population dynamics and diversity of Pseudomonas syringae on maple and pear trees and associated grasses. Phytopathology 78:1366–1370.CrossRefGoogle Scholar
  70. Maniatis, T., Fritsch E.F., and Sambrook, J. 1982. Molecular Cloning, a Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York. 545 pp.Google Scholar
  71. Martensson, A. and Gustafsson, J.G. 1985. Competition between Rhizobium trifolii strains for nodulation, during growth in a fermenter, and in soil-based inoculants, studied by ELISA. Journal of General Microbiology 131:3077–3082.Google Scholar
  72. Miller, L.T. 1982. Simple derivatization method for routine analysis of bacterial whole cell fatty acid methyl esters, including hydroxy acids. Journal of Clinical Microbiology 16:584–586.PubMedGoogle Scholar
  73. Morgan, J.A.W., Winstanley C., Pickup, R.W., Jones J.G., and Saunders, J.R. 1989. Direct phenotypic and genotypic detection of a recombinant pseudomonad population released into lake water. Applied and Environmental Microbiology 55:2537–2544.PubMedGoogle Scholar
  74. Moyed, H.S., Nguyen, T.T., and Bertrand, K.P. 1983. Multicopy Tn10 tet plasmids confer sensitivity to induction of tet gene expression. Journal of Bacteriology 155:549–556.PubMedGoogle Scholar
  75. Mullis, K.B. and Faloona, F.A. 1987. Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction. Methods in Enzymology 155:335–350.PubMedCrossRefGoogle Scholar
  76. Murray, R.E., Parsons, L.L., and Smith, M.S. 1990. Aerobic and anaerobic growth of rifampin-resistant denitrifying bacteria in soil. Applied and Environmental Microbiology 56:323–328.PubMedGoogle Scholar
  77. Muyzer, G. 1988. Applications of immunofluorescent techniques, pp. 219–222 in Sussman, M., Collins, C.H., Skinner F.A., and Stewart-Tull, D.E. (editors), The Release of Genetically-engineered Micro-organisms. Academic Press, London.Google Scholar
  78. Nakai, C., Hori, K., Kagamiyama, H., Nakazawa, T., and Nozaki, M. 1983. Purification, subunit structure, and partial amino acid sequence of metapyro-catechase. Journal of Biological Chemistry 258:2916–2922.PubMedGoogle Scholar
  79. Newell, S.Y. 1991. Estimating fungal biomass and productivity is decomposing litter. In Carroll, G.C. and Wicklow, D. (editors), The Fungal Community, 2nd ed. Dekker, New York. In press.Google Scholar
  80. Ngo, T.T. 1985. Ultrasensitive chromogen system for horse radish-peroxidase using 3-methyl-2-benzothiazolinone hydrazone and n-phenyldiethanolamine. Applied Biochemistry and Biotechnology 11:257–268.CrossRefGoogle Scholar
  81. Ogram, A., Sayler, G.S., and Barkay, T. 1988. DNA extraction and purification from sediments. Journal of Microbiological Methods 7:57–66.CrossRefGoogle Scholar
  82. Oliver, R.P., Roberts, I.N., Harling, R., Kenyon, L., Punt, P.J., Dingemanse, M.A., and van den Hondel, C.A.M.J.J. 1987. Transformation of Fulvia fulva, a fungal pathogen of tomato, to hygromycin B resistance. Current Genetics 12:231–233.CrossRefGoogle Scholar
  83. Ray, C., Igo, M., Shafer, W., Losick, R., and Moran, C.P., Jr. 1988. Suppression of etc promoter mutations in Bacillus subtilis. Journal of Bacteriology 170:900–907.PubMedGoogle Scholar
  84. Renwick, A. and Gareth-Jones, D. 1985. A comparison of the fluorescent ELISA and antibiotic resistance identification techniques for use in ecological experiments. Journal of Applied Bacteriology 58:199–206.Google Scholar
  85. Roberts, I.N., Oliver, R.P., Punt, P.J., and van den Hondel, C.A.M.J.J. 1989. Expression of the Escherichia coli β-glucuronidase gene in industrial and phytopathogenic filamentous fungi. Current Genetics 15:177–180.PubMedCrossRefGoogle Scholar
  86. Robinson, B.J., Pretzman, C.I., and Mattingly, J.A. 1983. Enzyme immunoassay in which a myeloma protein is used for detection of salmonellae. Applied and Environmental Microbiology 45:1816–1821.Google Scholar
  87. Rollins, D.M. and Colwell, R.R. 1986. Viable but nonculturable stage of Campylobacter jejuni and its role in survival in the natural aquatic environment. Applied and Environmental Microbiology 52:531–538.PubMedGoogle Scholar
  88. Rossman, A.Y., Palm, M.E., and Spielman, L.J. 1987. A Literature Guide for the Identification of Plant Pathogenic Fungi. American Phytopathological Society Press, St. Paul, MN. 252 pp.Google Scholar
  89. Roszak, D.B. and Colwell, R.R. 1987. Survival strategies of bacteria in the natural environment. Microbiological Reviews 51:365–379.PubMedGoogle Scholar
  90. Rovira, A.D., Newman, E.I., Bowen, H.J., and Campbell, R. 1974. Quantitative assessment of the rhizoplane microflora by direct microscopy. Soil Biology and Biochemistry 6:211–216.CrossRefGoogle Scholar
  91. Sala-Trepat, J.M. and Evans, W.C. 1971. The meta cleavage of catechol by Azotobacter species: 4-oxalocrotonate pathway. European Journal of Biochemistry 20:400–413.PubMedCrossRefGoogle Scholar
  92. Sasser, M. and Miller, L.T. 1984. Identification of pseudomonads by fatty acid profiling, pp. 45–46 in Psallidas, P.G. and Alivizatos, A.S. (editors), Proceedings of the Second Working Group on Pseudomonas syringae Pathovars. Hellenic Phytopathological Society Publishers, Athens, Greece.Google Scholar
  93. Schauer, A.T. 1988. Visualizing gene expression with luciferase fusions. Trends in Biotechnology 6:23–27.CrossRefGoogle Scholar
  94. Schmetterer, G., Wolk, C.P., and Elhai, J. 1986. Expression of luciferases from Vibrio harveyi and Vibrio fischeri in filamentous cyanobacteria. Journal of Bacteriology 167:411–414.PubMedGoogle Scholar
  95. Schroth, M.N., Thomson, S.V., and Moller, W.J. 1979. Streptomycin resistance in Envinia amylovora. Phytopathology 69:565–568.CrossRefGoogle Scholar
  96. Shaw, J.J. and Kado, C.I. 1986. Development of a Vibrio bioluminescence gene-set to monitor phytopathogenic bacteria during the ongoing disease process in a non-disruptive manner. Bio/Technology 4:560–564.CrossRefGoogle Scholar
  97. Shaw, J.J., Settles, L.G., and Kado, C.I. 1987. Transposon Tn4431 mutagenesis of Xanthomonas campestris pv. campestris: Characterization of a nonpathogenic mutant and cloning of a locus for pathogenicity. Molecular Plant-Microbe Interactions 1:39–45.CrossRefGoogle Scholar
  98. Southern, E.M. 1975. Detection of specific sequences among DNA fragments separated by gel electrophoresis. Journal of Molecular Biology 98:503–517.PubMedCrossRefGoogle Scholar
  99. Steffan, R.J. and Atlas, R.M. 1988. DNA amplification to enhance detection of genetically engineered bacteria in environmental samples. Applied and Environmental Microbiology 54:2185–2191.PubMedGoogle Scholar
  100. Steffan, R.J., Breen, A., Atlas, R.M., and Sayler, G.S. 1989. Monitoring genetically engineered microorganisms in freshwater microcosms. Journal of Industrial Microbiology 4:441–446.CrossRefGoogle Scholar
  101. Stotzky, G. and Babich, H. 1984. Fate of genetically engineered microbes in natural environments. Recombinant DNA Technical Bulletin 7:163–188.PubMedGoogle Scholar
  102. Torsvik, V.L. 1980. Isolation of bacterial DNA from soil. Soil Biology and Biochemistry 12:15–21.CrossRefGoogle Scholar
  103. Trevors, J.T., van Elsas, J.D., van Overbeek, L.S., and Starodub, M.-E. 1990. Transport of a genetically engineered Pseudomonas fluorescens strain through a soil microcosm. Applied and Environmental Microbiology 56:401–408.PubMedGoogle Scholar
  104. Vali, G. 1971. Quantitative evaluation of experimental results on the heterogeneous freezing nucleation of super cooled liquids. Journal of Atmospheric Science 28:402–409.CrossRefGoogle Scholar
  105. Van Gorcom, R.F.M., Punt, P.J., Pouwels, P.H., and van den Hondel, C. A.M.J.J. 1986. A system for the analysis of expression signals in Aspergillus. Gene 48:211–217.PubMedCrossRefGoogle Scholar
  106. Van Hartingsveldt, W., Mattern, I.E., van Zeijl, C.M.F., Pouwels P.H., and van den Hondel, C.A.M.J.J. 1987. Development of a homologous transformation system for Aspergillus niger based on the pyrG gene. Molecular and General Genetics 206:71–75.PubMedCrossRefGoogle Scholar
  107. Wall, L.E., Byers, D.M., and Meighen, E.A. 1984. In vivo and in vitro acylation of polypeptides in Vibrio harveyi: Identification of proteins involved in aldehyde production for bioluminescence. Journal of Bacteriology 159:720–724.PubMedGoogle Scholar
  108. Weller, D.M. and Saettler, A.W. 1978. Rifampin-resistant Xanthomonas phaseoli var. fuscans and Xanthomonas phaseoli: Tools for field study of bean blight bacteria. Phytopathology 68:778–781.CrossRefGoogle Scholar
  109. Winstanley, C., Morgan, J.A.W., Pickup, R.W., Jones, J.G., and Saunders, J.R. 1989. Differential regulation of lambda p l and p r promoters by a cI repressor in a broad-host-range thermoregulated plasmid marker system. Applied and Environmental Microbiology 65:771–777.Google Scholar
  110. Wright, S.F., Foster, J.G., and Bennett, O.L. 1986. Production and use of monoclonal antibodies for identification of strains of Rhizobium trifolii. Applied and Environmental Microbiology 52:119–123.PubMedGoogle Scholar
  111. Zimmermann, R. 1977. Estimation of bacterial number and biomass by epifluorescence microscopy and scanning microscopy, pp. 103–120 in Reinheimer, G. (editor), Microbial Ecology of a Brackish Water Environment. Springer-Verlag, Heidelberg, Federal Republic of Germany.Google Scholar
  112. Zola, H. 1987. Monoclonal Antibodies: A Manual of Techniques. CRC Press, Boca Raton, FL.Google Scholar
  113. Zukowski, M.M., Gaffney, D.F., Speck, D., Kauffmann, M., Findeli, A., Wisecup, A., and Lecocq, J. 1983. Chromogenic identification of genetic regulatory signals in Bacillus subtilis based on expression of a cloned Pseudomonas gene. Proceedings of the National Academy of Sciences USA 80:1101–1105.CrossRefGoogle Scholar

Copyright information

© Springer-Verlag New York Inc. 1991

Authors and Affiliations

  • David J. Drahos

There are no affiliations available

Personalised recommendations