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Appl. Environ. Microbiol., Sep 1996, 3333-3338, Vol 62, No. 9
Copyright © 1996, American Society for Microbiology

Recombinant Rhizobium meliloti strains with extra biotin synthesis capability

WR Streit and DA Phillips
Department of Agronomy and Range Science, University of California, Davis 95616, USA.

The growth of Rhizobium meliloti 1021 in an experimental alfalfa (Medicago sativa L.) rhizosphere was stimulated by adding nanomolar amounts of biotin. To overcome this biotin limitation, R. meliloti strains were constructed by conjugating the Escherichia coli biotin synthesis operon into biotin auxotroph R. meliloti 1021-B3. Transconjugant strains Rm1021-WS10 and Rm1021-WS11 grew faster in vitro and achieved a higher cell density than did R. meliloti 1021 and overproduced biotin on a defined medium. The increase in cell yield was associated with as much as a 99% loss in viability for Rm1021-WS11, but data suggested that a separate stabilizing factor in the E. coli DNA reduced cell death in Rm1021-WS10. In rhizosphere tests, the recombinant strains showed delayed growth and competed poorly against Rm1021.

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