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. 1990 May;56(5):1342–1346. doi: 10.1128/aem.56.5.1342-1346.1990

Oligonucleotide Probes That Hybridize with rRNA as a Tool To Study Frankia Strains in Root Nodules

Dittmar Hahn 1,*, Marjo J C Starrenburg 1, Antoon D L Akkermans 1
PMCID: PMC184406  PMID: 16348187

Abstract

Oligonucleotide probes that hybridize with specific sequences in variable regions of the 16S rRNA of the nitrogen-fixing actinomycete Frankia were used for the identification of Frankia strains in nodules. Frankia cells were released from plant tissue by grinding glutaraldehyde-fixed root nodules in guanidine hydrochloride solution. rRNA was obtained after sonication, precipitation with ethanol, and purification by phenolchloroform extraction. Degradation of rRNA, evident in Northern blots, did not affect hybridization with the oligonucleotides. Nodules of about 1 mg (fresh weight) provided sufficient rRNA for reliable detection of the Frankia strain. The utility of this rRNA extraction method was tested in a competition experiment between two effective Frankia strains on cloned Alnus glutinosa plants.

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Selected References

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  1. Church G. M., Gilbert W. Genomic sequencing. Proc Natl Acad Sci U S A. 1984 Apr;81(7):1991–1995. doi: 10.1073/pnas.81.7.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Embley T. M., Smida J., Stackebrandt E. Reverse transcriptase sequencing of 16S ribosomal RNA from Faenia rectivirgula, Pseudonocardia thermophila and Saccharopolyspora hirsuta, three wall type IV actinomycetes which lack mycolic acids. J Gen Microbiol. 1988 Apr;134(4):961–966. doi: 10.1099/00221287-134-4-961. [DOI] [PubMed] [Google Scholar]
  3. Giovannoni S. J., DeLong E. F., Olsen G. J., Pace N. R. Phylogenetic group-specific oligodeoxynucleotide probes for identification of single microbial cells. J Bacteriol. 1988 Feb;170(2):720–726. doi: 10.1128/jb.170.2.720-726.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Govers F., Gloudemans T., Moerman M., van Kammen A., Bisseling T. Expression of plant genes during the development of pea root nodules. EMBO J. 1985 Apr;4(4):861–867. doi: 10.1002/j.1460-2075.1985.tb03711.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Göbel U. B., Geiser A., Stanbridge E. J. Oligonucleotide probes complementary to variable regions of ribosomal RNA discriminate between Mycoplasma species. J Gen Microbiol. 1987 Jul;133(7):1969–1974. doi: 10.1099/00221287-133-7-1969. [DOI] [PubMed] [Google Scholar]
  6. Lane D. J., Pace B., Olsen G. J., Stahl D. A., Sogin M. L., Pace N. R. Rapid determination of 16S ribosomal RNA sequences for phylogenetic analyses. Proc Natl Acad Sci U S A. 1985 Oct;82(20):6955–6959. doi: 10.1073/pnas.82.20.6955. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Normand P., Simonet P., Bardin R. Conservation of nif sequences in Frankia. Mol Gen Genet. 1988 Aug;213(2-3):238–246. doi: 10.1007/BF00339587. [DOI] [PubMed] [Google Scholar]
  8. Olsen G. J., Lane D. J., Giovannoni S. J., Pace N. R., Stahl D. A. Microbial ecology and evolution: a ribosomal RNA approach. Annu Rev Microbiol. 1986;40:337–365. doi: 10.1146/annurev.mi.40.100186.002005. [DOI] [PubMed] [Google Scholar]
  9. Simonet P., Le N. T., Du Cros E. T., Bardin R. Identification of frankia strains by direct DNA hybridization of crushed nodules. Appl Environ Microbiol. 1988 Oct;54(10):2500–2503. doi: 10.1128/aem.54.10.2500-2503.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Stahl D. A., Flesher B., Mansfield H. R., Montgomery L. Use of phylogenetically based hybridization probes for studies of ruminal microbial ecology. Appl Environ Microbiol. 1988 May;54(5):1079–1084. doi: 10.1128/aem.54.5.1079-1084.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Tenover F. C. Diagnostic deoxyribonucleic acid probes for infectious diseases. Clin Microbiol Rev. 1988 Jan;1(1):82–101. doi: 10.1128/cmr.1.1.82. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Verwoerd T. C., Dekker B. M., Hoekema A. A small-scale procedure for the rapid isolation of plant RNAs. Nucleic Acids Res. 1989 Mar 25;17(6):2362–2362. doi: 10.1093/nar/17.6.2362. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Viscidi R. P., Yolken R. G. Molecular diagnosis of infectious diseases by nucleic acid hybridization. Mol Cell Probes. 1987 Mar;1(1):3–14. doi: 10.1016/0890-8508(87)90003-x. [DOI] [PubMed] [Google Scholar]

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