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Mutational analysis of the Bradyrhizobium japonicum common nod genes and further nod box-linked genomic DNA regions

Molecular and General Genetics MGG volume 215, pages 407–415 (1989)Cite this article

Summary

By insertional and deletional marker replacement mutagenesis the common nod region of Bradyrhizobium japonicum was examined for the presence of additional, essential nodulation genes. An open reading frame located in the 800 bp large intergenic region between nodD1 and nodA did not appear to be essential for nodulation of soybean. Furthermore, a strain with a deletion of the nodI- and nodJ-like genes downstream of nodC had a Nod+ phenotype. A mutant with a 1.7 kb deletion immediately downstream of nodD1 considerably delayed the onset of nodulation. This region carried a second copy of nodD (nodD2). A nodD1-nodD2 double mutant had a similar phenotype to the nodD2 mutant. Using a 22-mer oligonucleotide probe partially identical to the nod box sequence, a total of six hybridizing regions were identified in B. japonicum genomic DNA and isolated from a cosmid library. Sequencing of the hybridizing regions revealed that at least three of them represented true nod box sequences whereas the others showed considerable deviations from the consensus sequence. One of the three nod box sequences was the one known to be associated with nodA, whereas the other two were located 60 to 70 kb away from nif cluster I. A deletion of one of these two sequences plus adjacent DNA material mmutant Δ308) led to a reduced nodulation on Vigna radiata but not on soybean. Thus, this region is probably involved in the determination of host specificity.

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Affiliations

  1. ETH-Zentrum, Mikrobiologisches Institut, Eidgenössische Technische Hochschule, Schmelzbergstrasse 7, CH-8092, Zürich, Switzerland

    Michael Göttfert, Joseph W. Lamb, Regula Gasser, Jan Semenza & Hauke Hennecke

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  1. Michael Göttfert
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  2. Joseph W. Lamb
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  3. Regula Gasser
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  4. Jan Semenza
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  5. Hauke Hennecke
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Dedicated to Prof. Giorgio Semenza on the occasion of his 60th birthday

Communicated by J. Lengeler

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Göttfert, M., Lamb, J.W., Gasser, R. et al. Mutational analysis of the Bradyrhizobium japonicum common nod genes and further nod box-linked genomic DNA regions. Molec. Gen. Genet. 215, 407–415 (1989). https://doi.org/10.1007/BF00427037

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  • DOI: https://doi.org/10.1007/BF00427037

Key words

  • Bradyrhizobium
  • Nodulation
  • nod genes
  • Soybean
  • Symbiosis