Plant and Soil

, Volume 194, Issue 1–2, pp 25–36 | Cite as

Isolation of endophytic bacteria from rice and assessment of their potential for supplying rice with biologically fixed nitrogen

  • J.R. Stoltzfus
  • R. So
  • P.P. Malarvithi
  • J.K. Ladha
  • F.J. de Bruijn


The extension of nitrogen-fixing symbioses to important crop plants such as the cereals has been a long-standing goal in the field of biological nitrogen fixation. One of the approaches that has been used to try to achieve this goal involves the isolation and characterization of stable endophytic bacteria from a variety of wild and cultivated rice species that either have a natural ability to fix nitrogen or can be engineered to do so. Here we present the results of our first screening effort for rice endophytes and their characterization using acetylene reduction assays (ARA), genomic fingerprinting with primers corresponding to naturally occurring repetitive DNA elements (rep-PCR), partial 16S rDNA sequence analysis and PCR mediated detection of nitrogen fixation (nif) genes with universal nif primers developed in our laboratory. We also describe our efforts to inoculate rice plants with the isolates obtained from the screening, in order to examine their invasiveness and persistence (stable endophytic maintenance). Lastly, we review our attempts to tag selected isolates with reporter genes/proteins, such as beta-glucuronidase (gus) or green fluorescent protein (gfp), in order to be able to track putative endophytes during colonization of rice tissues.

endophytic microbes endo-symbiosis genomic fingerprinting infection marker genes nitrogen fixation PCR rep-PCR rice 


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Copyright information

© Kluwer Academic Publishers 1997

Authors and Affiliations

  • J.R. Stoltzfus
    • 1
  • R. So
    • 2
  • P.P. Malarvithi
    • 2
  • J.K. Ladha
    • 2
  • F.J. de Bruijn
    • 1
    • 3
    • 4
  1. 1.MSU-DOE Plant Research LaboratoryThe Philippines
  2. 2.International Rice Research InstituteLos BanosThe Philippines
  3. 3.Department of MicrobiologyThe Philippines
  4. 4.NSF Center for Microbial EcologyMichigan State UniversityE. LansingU.S.A

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