A molecular linkage map of nitrogenase and nodulation genes in Rhizobium trifolii

Peter R. Schofield; Michael A. Djordjevic; Barry G. Rolfe; John Shine; John M. Watson

doi:10.1007/BF00392191

A molecular linkage map of nitrogenase and nodulation genes in Rhizobium trifolii

Peter R. Schofield, Michael A. Djordjevic, Barry G. Rolfe, John Shine, John M. Watson^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

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Abstract

A molecular map was constructed linking the nitrogenase structural genes (nif) and nodulation genes (nod) in the white clover symbiont, Rhizobium trifolii. In R. trifolii strain ANU843 these two genetic regions are located some 16 kilobases (kb) apart on the 180 kb symbiotic (Sym) plasmid. The molecular linkage of nod and nif genetic regions was established by hybridization analysis using recombinant plasmids containing overlapping cloned sequences. Nodulation genes were located by means of a Tn5-induced nodulation-defective mutant that failed to induce clover root hair curling (Hac^- phenotype). A cloned wild-type DNA fragment was shown to phenotypically correct the Hac^- mutation by complementation. The nifHDK genes were cloned by positive hybridization to another R. trifolii nif-specific probe. Location of the nif genes relative to the nod genes was established by analysis of a Sym plasmid deletion derivative.

Original language	English
Pages (from-to)	459-465
Number of pages	7
Journal	MGG Molecular & General Genetics
Volume	192
Issue number	3
DOIs	https://doi.org/10.1007/BF00392191
Publication status	Published - Nov 1983

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title = "A molecular linkage map of nitrogenase and nodulation genes in Rhizobium trifolii",

abstract = "A molecular map was constructed linking the nitrogenase structural genes (nif) and nodulation genes (nod) in the white clover symbiont, Rhizobium trifolii. In R. trifolii strain ANU843 these two genetic regions are located some 16 kilobases (kb) apart on the 180 kb symbiotic (Sym) plasmid. The molecular linkage of nod and nif genetic regions was established by hybridization analysis using recombinant plasmids containing overlapping cloned sequences. Nodulation genes were located by means of a Tn5-induced nodulation-defective mutant that failed to induce clover root hair curling (Hac- phenotype). A cloned wild-type DNA fragment was shown to phenotypically correct the Hac- mutation by complementation. The nifHDK genes were cloned by positive hybridization to another R. trifolii nif-specific probe. Location of the nif genes relative to the nod genes was established by analysis of a Sym plasmid deletion derivative.",

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T1 - A molecular linkage map of nitrogenase and nodulation genes in Rhizobium trifolii

AU - Schofield, Peter R.

AU - Djordjevic, Michael A.

AU - Rolfe, Barry G.

AU - Shine, John

AU - Watson, John M.

PY - 1983/11

Y1 - 1983/11

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AB - A molecular map was constructed linking the nitrogenase structural genes (nif) and nodulation genes (nod) in the white clover symbiont, Rhizobium trifolii. In R. trifolii strain ANU843 these two genetic regions are located some 16 kilobases (kb) apart on the 180 kb symbiotic (Sym) plasmid. The molecular linkage of nod and nif genetic regions was established by hybridization analysis using recombinant plasmids containing overlapping cloned sequences. Nodulation genes were located by means of a Tn5-induced nodulation-defective mutant that failed to induce clover root hair curling (Hac- phenotype). A cloned wild-type DNA fragment was shown to phenotypically correct the Hac- mutation by complementation. The nifHDK genes were cloned by positive hybridization to another R. trifolii nif-specific probe. Location of the nif genes relative to the nod genes was established by analysis of a Sym plasmid deletion derivative.

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A molecular linkage map of nitrogenase and nodulation genes in Rhizobium trifolii

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