Antonie van Leeuwenhoek

, Volume 105, Issue 2, pp 389–399 | Cite as

Genetic diversity of rhizobia isolated from nodules of the relic species Vavilovia formosa (Stev.) Fed.

  • Vera I. Safronova
  • Anastasiia K. Kimeklis
  • Elena P. Chizhevskaya
  • Andrey A. Belimov
  • Evgeny E. Andronov
  • Alexander G. Pinaev
  • Andrey R. Pukhaev
  • Konstantin P. Popov
  • Igor A. Tikhonovich
Original Paper

Abstract

Sixteen bacterial strains were isolated from root nodules of Vavilovia formosa plants originated from the North Ossetian State Natural Reserve (Caucasus, Russia). Phylogenetic analysis of these strains was performed using partial 16S rRNA gene and internally transcribed spacer (ITS) sequences. The results showed that the isolates belong to three families of root nodule bacteria. Twelve of them were related to the genus Rhizobium (family Rhizobiaceae) but four strains can be most probably identified as Phyllobacterium-related (family Phyllobacteriaceae), Bosea- and Rhodopseudomonas-related (family Bradyrhizobiaceae). Amplified fragment length polymorphism clustering was congruent with ITS phylogeny but displayed more variability for Rhizobium isolates, which formed a single group at the level of 30 % similarity. We expect that the isolates obtained can belong to new taxa at genus, species or subspecies levels. The results of PCR amplification of the nodulation genes nodC and nodX showed their presence in all Rhizobium isolates and one Rhodopseudomonas-related isolate. The nodC gene sequences of V. formosa isolates were closely related to those of the species Rhizobium leguminosarum bv. viciae but formed separate clusters and did not intermingle with any reference strains. The presence of the nodX gene, which is necessary for nodulation of Afghan peas (Pisum sativum L.) originated from the Middle East, allows the speculation that these wild-type pea cultivars may be the closest existing relatives of V. formosa. Thus, the studies of genetic diversity and symbiotic genes of V. formosa microsymbionts provide the primary information about their phylogeny and contribute to the conservation of this relict leguminous species.

Keywords

Vavilovia formosa (Stev.) Fed. Root nodule bacteria 16S and ITS rDNA sequencing AFLP fingerprinting Rhizobial biodiversity 

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Copyright information

© Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  • Vera I. Safronova
    • 1
  • Anastasiia K. Kimeklis
    • 1
  • Elena P. Chizhevskaya
    • 1
  • Andrey A. Belimov
    • 1
  • Evgeny E. Andronov
    • 1
  • Alexander G. Pinaev
    • 1
  • Andrey R. Pukhaev
    • 2
  • Konstantin P. Popov
    • 3
  • Igor A. Tikhonovich
    • 1
  1. 1.All-Russia Research Institute for Agricultural Microbiology (ARRIAM)Saint PetersburgRussia
  2. 2.Gorsky State Agrarian UniversityVladikavkazRussia
  3. 3.North-Ossetian Nature Reserve, Republic North-Ossetia-AlaniaAlagirRussia

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