Genetic complementation of rhizobial nod mutants with Frankia DNA:: artifact or reality?

Two divergent reports have been published on the genetic complementation of rhizobial nod mutants using Frankia DNA. In 1991 putative Frankia cosmid library clones were reported to restore normal nodulation properties to Rhizobium leguminosarum biovar vicine nodD::Tn5, but no supporting sequence data were published. In 1992 a second group reported a failure to find any evidence of functional complementation of Various rhizobial nod mutants by Frankia DNA (nodA, nodB and nodC). Complementation tests of nine Nod(-) R. leguminosarum by. viciae or Sinorhizobium meliloti Tn5 mutants (nodA(-), nodB(-), nodC(-), nodD(-), nodF(-), nodL(-), nodH(-)) were thus performed using a Frankia gene library in pLAFR3 to clarify this situation. Rhizobial transconjugants obtained by tri-parental matings were screened for restoration of the nodulation phenotype on their host plants, Vicia sativa subsp. nigra or Medicago sativa. Nodulation was observed on plants inoculated with transconjugants of the R. leguminosarum by. viciae nodC::Tn5 mutant. The Nod(+) rhizobial transconjugants were isolated and analysed. The Nod(+) phenotype of these transconjugants was found to be due to Tn5 excision/transposition. No functional complementation was found with any of the mutants used, suggesting that rhizobial complementation of nod mutants with Frankia DNA is unlikely to occur.